Pyridinone antagonists of alpha-4 integrins

ABSTRACT

The present invention provides compounds that are alpha4 integrin antagonists having a structure according to the following formula: 
     
       
         
         
             
             
         
       
     
     or a tautomer, mixture of tautomers, salt or solvate thereof, wherein Cy, ring A, m, n, p, R 1 , R 2 , R 3 , R 4 , R 5  and R 6  are defined in the specification. The invention further provides pharmaceutical compositions including the compounds of the invention as well as methods of making and using the compounds and compositions of the invention, e.g., in the treatment and prevention of various conditions and disorders, such as Crohn&#39;s disease and ulcerative colitis.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Continuation Application of U.S. patentapplication Ser. No. 12/768,680, filed Apr. 27, 2010, now U.S. Pat. No.8,367,836, which claims the benefit under 35 U.S.C. §119 (e) of U.S.Provisional Application Ser. No. 61/172,876, filed Apr. 27, 2009, bothof which are hereby incorporated by reference in their entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to compounds which inhibit leukocyte adhesionand, in particular, leukocyte adhesion mediated by α₄β₇. Accordingly,compounds of this invention are useful in the treatment and preventionof diseases mediated by α₄β₇ binding and cell adhesion and activationsuch as multiple sclerosis, asthma, allergic rhinitis, rheumatoidarthritis, septic arthritis, restenosis, inflammatory bowel diseaseincluding ulcerative colitis and Crohn's disease, dermatitis, psoriasis,and the like.

2. State of the Art

The following publications, patents and patent applications are cited inthis application as superscript numbers:

-   Tidswell, et al., J. of Immunology, 1497-1505 (1997)-   Springer, Nature, 346:425-434 (1990)-   Osborn, Cell, 62:3-6 (1990)-   Vedder, et al., Surgery, 106:509 (1989)-   Pretolani, et al., J. Exp. Med., 180:795 (1994)-   Abraham. et al., J. Clin. Invest., 93:776 (1994)-   Mulligan, et al., J. Immunology, 150:2407 (1993)-   Cybulsky, et al., Science, 251:788 (1991)-   Li, et al., Arterioscler. Thromb., 13:197 (1993)-   Sasseville, et al., Am. J. Path., 144:27 (1994)-   Yang, et al., Proc. Nat. Acad. Science (USA), 90:10494 (1993)-   Burkly, et al., Diabetes, 43:529 (1994)-   Baron, et al., J. Clin. Invest., 93:1700 (1994)-   Hamann, et al., J. Immunology, 152:3238 (1994)-   Yednock, et al., Nature, 356:63 (1992)-   Baron, et al., J. Exp. Med., 177:57 (1993)-   van Dinther-Janssen, et al., J. Immunology, 147:4207 (1991)-   van Dinther-Janssen, et al., Annals. Rheumatic Dis., 52:672 (1993)-   Elices, et al, J. Clin. Invest., 93:405 (1994)-   Postigo, et al., J. Clin. Invest., 89:1445 (1991)-   Paul, et al., Transpl. Proceed, 25:813 (1993)-   Okarhara, et al., Can. Res., 54:3233 (1994)-   Paavonen, et al., Int. J. Can., 58:298 (1994)-   Schadendorf et al., J. Path., 170:429 (1993)-   Bao, et al., Diff., 52:239 (1993)-   Lauri, et al., British J. Cancer, 68:862 (1993)-   Kawaguchi, et al., Japanese J. Cancer Res., 83:1304 (1992)-   Kogan, et al., U.S. Pat. No. 5,510,332, issued Apr. 23, 1996-   International Patent Appl. Publication No. WO 96/01644

All of the above publications, patents and patent applications areherein incorporated by reference in their entirety to the same extent asif each individual publication, patent or patent application wasspecifically and individually indicated to be incorporated by referencein its entirety.

Integrins are heterodimeric adhesion receptors that mediate cell-celland cell-extracellular matrix interactions. The β₇ integrin subfamilyhas two known members: α₄β₇ and α_(E)β₇. These β₇ integrins areexpressed primarily by leukocytes. β₇ integrins are unique among knownintegrins in their ability to recognize certain ligands expressed on thesurface of endothelial and epithelial cells in mucosal organs.¹

α₄β₇ is a lymphocyte homing receptor and plays a crucial role in themigration of these cells to the intestine and associated lymphoidtissue, such as Peyer's patches in the intestine. α₄β₇ mediates adhesionto a ligand on Peyer's patch high endothelial venules (“HEV⁴”). Theligand on Peyer's patch HEV is MAdCAM-1, a glycoprotein in the Igsuperfamily. MAdCAM-1 is expressed on Peyer's patch HEV, mesentericlymph node HEV, and lamina propria venules within the gut. Antibodiesagainst α₄ or β₇ subunits inhibit attachment of circulating lymphocytesto Peyer's patch HEV in vivo.¹

Memory T cells that circulate preferentially to intestinal tissuesexpress high levels of α₄β₇, whereas those that circulate to otherorgans are mostly α₄β₁. These α₄β₇ memory T cells express a relatedintegrin, α₄β₁, which is not able to mediate cell adhesion to MAdCAM-1.However, both α₄β₇ and α₄β₁, can mediate adhesion to VCAM-1 and tofibronectin.¹

Intercellular adhesion mediated by α₄β₇ and other cell surface receptorsis associated with a number of inflammatory responses. At the site of aninjury or other inflammatory stimulus, activated vascular endothelialcells express molecules that are adhesive for leukocytes. The mechanicsof leukocyte adhesion to endothelial cells involves, in part, therecognition and binding of cell surface receptors on leukocytes to thecorresponding cell surface molecules on endothelial cells. Once bound,the leukocytes migrate across the blood vessel wall to enter the injuredsite and release chemical mediators to combat infection. For reviews ofadhesion receptors of the immune system, see, for example, Springer² andOsborn.³

Inflammatory brain disorders, such as experimental autoimmuneencephalomyelitis (EAE), multiple sclerosis (MS) and meningitis, areexamples of central nervous system disorders in which theendothelium/leukocyte adhesion mechanism results in destruction tootherwise healthy brain tissue. Large numbers of leukocytes migrateacross the blood brain barrier (BBB) in subjects with these inflammatorydiseases. The leukocytes release toxic mediators that cause extensivetissue damage resulting in impaired nerve conduction and paralysis.

In other organ systems, tissue damage also occurs via an adhesionmechanism resulting in migration or activation of leukocytes. Forexample, it has been shown that the initial insult following myocardialischemia to heart tissue can be further complicated by leukocyte entryto the injured tissue causing still further insult.⁴ Other inflammatoryconditions mediated by an adhesion mechanism include, by way of example,asthma,⁵⁻⁷ Alzheimer's disease, atherosclerosis,⁸⁻⁹ AIDS dementia,¹⁰diabetes (including acute juvenile onset diabetes),¹¹⁻¹³ inflammatorybowel disease (including ulcerative colitis and Crohn's disease),⁴multiple sclerosis,¹⁵⁻¹⁶ rheumatoid arthritis,¹⁷⁻²⁰ tissuetransplantation,²¹ tumor metastasis,²²⁻²⁷ meningitis, encephalitis,stroke, and other cerebral traumas, nephritis, retinitis, atopicdermatitis, psoriasis, myocardial ischemia and acute leukocyte-mediatedlung injury such as that which occurs in adult respiratory distresssyndrome.

Despite advances in the understanding of leukocyte adhesion, the art hasonly recently addressed the use of inhibitors of adhesion in thetreatment of inflammatory conditions.^(28,29) Novel alpha4 antagonistsare needed for the development of additional treatment options forinflammatory diseases, such as MS, asthma, Crohn's disease andrheumatoid arthritis. Furthermore, assays useful for the diagnosis ofα₄β₇ mediated conditions are needed. The current invention addressesthese and other needs.

SUMMARY OF THE INVENTION

In various aspects, the invention provides compounds of Formula (Ia):

or a salt or solvate or single stereoisomer or mixture of stereoisomersthereof,wherein

-   -   m is an integer selected from 0 to 4;    -   n is an integer selected from 0 to 3;    -   p is an integer selected from 0 to 4;    -   ring A is a member selected from substituted or unsubstituted        aryl and substituted or unsubstituted heteroaryl;    -   Cy is a member selected from substituted or unsubstituted        (C₃-C₁₀)cycloalkyl, substituted or unsubstituted 3- to        10-membered heterocycloalkyl, substituted or unsubstituted aryl        and substituted or unsubstituted heteroaryl;    -   each R¹ and each R² is a member independently selected from H,        substituted or unsubstituted (C₁-C₁₀)alkyl, substituted or        unsubstituted 2- to 10-membered heteroalkyl, substituted or        unsubstituted (C₃-C₁₀)cycloalkyl, substituted or unsubstituted        3- to 10-membered heterocycloalkyl, substituted or unsubstituted        aryl, substituted or unsubstituted heteroaryl, nitro, CN,        halogen, OR¹², SR¹², NR¹²R¹³, C(O)R¹⁴, C(O)NR¹²R¹³,        OC(O)NR¹²R¹³, C(O)OR¹², NR¹⁵C(O)R¹⁴, NR¹⁵C(O)OR¹²,        NR¹⁵C(O)NR¹²R¹³, NR¹⁵C(S)NR¹²R¹³, NR¹⁵S(O)₂R¹⁴, S(O)₂NR¹²R¹³ and        S(O)_(z)R¹⁴,    -   wherein        -   z is 1 or 2;        -   R¹², R¹³ and R¹⁵ are members independently selected from H,            acyl, substituted or unsubstituted (C₁-C₁₀)alkyl,            substituted or unsubstituted 2- to 10-membered heteroalkyl,            substituted or unsubstituted aryl, substituted or            unsubstituted heteroaryl, substituted or unsubstituted            (C₃-C₁₀)cycloalkyl, substituted or unsubstituted 3- to            10-membered heterocycloalkyl; and        -   R¹⁴ is a member independently selected from substituted or            unsubstituted (C₁-C₁₀)alkyl, substituted or unsubstituted 2-            to 10-membered heteroalkyl, substituted or unsubstituted            aryl, substituted or unsubstituted heteroaryl, substituted            or unsubstituted (C₃-C₁₀)cycloalkyl, substituted or            unsubstituted 3- to 10-membered heterocycloalkyl,        -   wherein R¹² and R¹³, together with the nitrogen atoms to            which they are attached, are optionally joined to form a 4-            to 7-membered ring, and        -   wherein two adjacent R¹, together with the atoms to which            they are attached, are optionally joined to form a 5- to            7-membered ring, and        -   wherein two adjacent R², together with the atoms to which            they are attached, are optionally joined to form a 5- to            7-membered ring;    -   R³, R⁴ and R⁵ are members independently selected from H,        substituted or unsubstituted (C₁-C₄)alkyl, halogen and CN; and        R⁶ is a member selected from H and substituted or unsubstituted        (C₁-C₄)alkyl.

The invention is also directed to a compound of Formula (Ia′), Formula(Ib), or Formula (Ic)

-   -   wherein Cy, R¹, R², R³, R⁴, R⁵, R⁶, n, p, m, and A are as        defined as for Formula (Ia);    -   m1 is an integer selected from 0 to 3;    -   R^(1a) and R^(1b) are independently selected from the same group        as for R¹ and R² as defined above.

Also included in the invention is a salt or solvate or singlestereoisomer or mixture of stereoisomers thereof, wherein m is aninteger selected from 0 to 4, n is an integer selected from 0 to 4, p isan integer selected from 0 to 6 and m1 is an integer selected from 0 to3. In one example, n is selected from 1 to 4. In another example, n isselected from 1 and 2. In yet another example, n is 1. In a furtherexample, m is 0 or 1. In another example, p is 0 or 1.

In Formula (Ia), Formula (Ia′), Formula (Ib) or Formula (Ic), ring A isa member selected from substituted or unsubstituted aryl (e.g., phenyl,naphthyl) and substituted or unsubstituted heteroaryl (e.g., pyridyl,thiophene, thiazole, imidazolyl).

In Formula (Ia), Formula (Ia′), Formula (Ib) or Formula (Ic), Cy is amember selected from substituted or unsubstituted cycloalkyl (e.g.,C₃-C₁₀ cycloalkyl), substituted or unsubstituted heterocycloalkyl (e.g.,3- to 10-membered heterocycloalkyl), substituted or unsubstituted aryl(e.g., phenyl or naphthyl) and substituted or unsubstituted heteroaryl(e.g., pyridyl). In one example in the above structures, Cy issubstituted or unsubstituted aryl (e.g., phenyl or naphthyl) orsubstituted or unsubstituted heteroaryl (e.g., pyridyl). In a particularexample, Cy in the above structures is a member selected fromsubstituted or unsubstituted phenyl (e.g., substituted or unsubstituted4-chloro-phenyl or substituted or unsubstituted 4-phenoxyphenyl),substituted or unsubstituted naphthyl, substituted or unsubstitutedbiphenyl (e.g., substituted or unsubstituted biphenyl-4-yl) andsubstituted or unsubstituted pyridyl (e.g., substituted or unsubstituted4-pyridyl).

In Formula (Ia), Formula (Ia′), Formula (Ib) or Formula (Ic), each R¹,each R², R^(1a) and R^(1b) are members independently selected from H,substituted or unsubstituted alkyl (e.g., C₁-C₁₀ alkyl), substituted orunsubstituted heteroalkyl (e.g., 2- to 10-membered heteroalkyl),substituted or unsubstituted cycloalkyl (e.g., C₃-C₁₀ cycloalkyl),substituted or unsubstituted heterocycloalkyl (e.g., 3- to 10-memberedheterocycloalkyl), substituted or unsubstituted aryl (e.g., substitutedor unsubstituted phenyl or naphthyl), substituted or unsubstitutedheteroaryl (e.g., substituted or unsubstituted pyridyl), nitro, CN,halogen (e.g., I, F, Cl or Br), OR¹², SR¹², NR¹²R¹³, C(O)R¹⁴,C(O)NR¹²R¹³, OC(O)NR¹²R¹³, C(O)OR¹², NR¹⁵C(O)R¹⁴, NR¹⁵C(O)OR¹²,NR¹⁵C(O)NR¹²R¹³, NR¹⁵C(S)NR¹²R¹³, NR¹⁵S(O)₂R¹⁴, S(O)₂NR¹²R¹³ andS(O)_(z)R¹⁴, wherein z is 1 or 2. R¹², R¹³ and R¹⁵ are membersindependently selected from H, acyl, substituted or unsubstituted alkyl(e.g., C₁-C₁₀ alkyl), substituted or unsubstituted heteroalkyl (e.g., 2-to 10-membered heteroalkyl), substituted or unsubstituted aryl,substituted or unsubstituted heteroaryl, substituted or unsubstitutedcycloalkyl (e.g., C₃-C₁₀ cycloalkyl) and substituted or unsubstitutedheterocycloalkyl (e.g., 3- to 10-membered heterocycloalkyl). R¹⁴ is amember independently selected from substituted or unsubstituted alkyl(e.g., C₁-C₁₀ alkyl), substituted or unsubstituted heteroalkyl (e.g., 2-to 10-membered heteroalkyl), substituted or unsubstituted aryl,substituted or unsubstituted heteroaryl, substituted or unsubstitutedcycloalkyl (e.g., C₃-C₁₀ cycloalkyl) and substituted or unsubstitutedheterocycloalkyl (e.g., 3- to 10-membered heterocycloalkyl). R¹² andR¹³, together with the nitrogen atoms to which they are attached, areoptionally joined to form a 4- to 7-membered ring. Two adjacent R¹,together with the atoms to which they are attached, are optionallyjoined to form a 5- to 7-membered ring. Two adjacent R², together withthe atoms to which they are attached, are optionally joined to form a 5-to 7-membered ring.

In Formula (Ia), Formula (Ia′), Formula (Ib) or Formula (Ic), R³, R⁴ andR⁵ are members independently selected from H, substituted orunsubstituted alkyl (e.g., C₁-C₄ alkyl), halogen (e.g., I, F, Cl, Br),substituted or unsubstituted aryl, —O-aryl, and CN. In a particularexample in the above structures, R³, R⁴ and R⁵ are each H.

In Formula (Ia), Formula (Ia′), Formula (Ib) or Formula (Ic), R⁶ is amember selected from H, substituted or unsubstituted alkyl (e.g., C₁-C₆alkyl), substituted or unsubstituted heteroalkyl (e.g., 2- to 6-memberedheteroalkyl), substituted or unsubstituted cycloalkyl (e.g., C₃-C₈cycloalkyl), substituted or unsubstituted heterocycloalkyl (e.g., 3- to8-membered heterocycloalkyl), substituted or unsubstituted aryl (e.g.,substituted or unsubstituted phenyl), substituted or unsubstitutedheteroaryl (e.g., substituted or unsubstituted pyridyl), NR¹²R¹³,C(O)R¹⁴, (C(O)NR¹²R¹³, C(O)OR¹² and S(O)_(z)R¹⁴, wherein z is 1 or 2 andR¹², R¹³ and R¹⁴ are defined as herein above. In one example, R⁶ is amember selected from H and substituted or unsubstituted alkyl (e.g.,C₁-C₄ alkyl). In a particular example in the above structures, R⁶ is H.

In Formula (Ic), E is selected from O, S, C(O), S(O)₂ and NR⁴⁰, whereinR⁴⁰ is a member selected from substituted or unsubstituted alkyl (e.g.,C₁-C₁₀ alkyl). In one example, R⁴⁰ is unsubstituted C₁-C₄ alkyl (e.g.,methyl or ethyl).

In one embodiment, the invention is directed to compounds of FIG. 1and/or FIG. 2 or salts or solvates thereof.

In another embodiment is provided a pharmaceutical compositioncomprising a compound according to any one of the preceding claims and apharmaceutically acceptable carrier.

In another embodiment, the invention is directed to a method of treatingan inflammatory disease comprising administering to a mammalian subjectin need thereof a pharmaceutically effective amount of a compound of theinvention. In one embodiment, the inflammatory disease is a memberselected from asthma, inflammatory bowel disease, ulcerative colitis,Crohn's disease, multiple sclerosis, rheumatoid arthritis, tumormetastasis, graft versus host disease, and organ or tissue rejection. Inone embodiment the disease is ulcerative colitis or Crohn's disease.

In yet another embodiment, the invention is directed to use of acompound of the invention in an in vitro assay measuring binding of anα4β1 or α4β7 integrin to an integrin ligand. In one embodiment, theintegrin ligand is a member selected from fibronectin (FN), VCAM-1,osteopontin and MadCAM.

In one embodiment, the assay comprises:

-   -   (i) binding the ligand to a surface;    -   (ii) contacting the ligand with a cell expressing the integrin,        in the presence of the compound; and    -   (iii) measuring the amount of cells bound to the surface.

In another embodiment, the invention is directed to use of a compound ofthe invention in an in vitro assay measuring binding of the compound toan α4β1 or α4β7 integrin in the presence of a candidate molecule. Inanother embodiment, the invention is directed to use of a compound ofthe invention in an in vitro assay for identifying a candidate moleculecapable of binding to α4β1 or α4β7 integrin. In one embodiment, theassay is a competitive binding assay.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1I show a table summarizing exemplary compounds of theinvention (compounds 1-121) and their in vitro biological activities.The MadCAM Adhesion Assay (A) and the MadCAM SRU Adhesion Assay (B),which were used to generate the presented data, are described herein inExamples 32 and 33, respectively.

FIGS. 2A-2Z, 2AA-2ZZ, 2AAA and 2BBB show a table summarizing exemplarycompounds of the invention and their in vitro biological activities. TheMadCAM Adhesion Assay (A) and the MadCAM SRU Adhesion Assay (B), whichwere used to generate the presented data, are described herein inExamples 32 and 33, respectively.

DETAILED DESCRIPTION OF THE INVENTION Definitions

The definitions and explanations below are for the terms as usedthroughout this entire document including both the specification and theclaims. Throughout the specification and the appended claims, a givenformula or name shall encompass all isomers thereof, such asstereoisomers, geometrical isomers, optical isomers, tautomers, andmixtures thereof where such isomers exist, as well as pharmaceuticallyacceptable salts and solvates thereof, such as hydrates.

It should be noted that, as used in this specification and the appendedclaims, the singular forms “a,” “an,” and “the” include plural referentsunless the content clearly dictates otherwise. Thus, for example,reference to a composition containing “a compound” includes a mixture oftwo or more compounds. It should also be noted that the term “or” isgenerally employed in its sense including “and/or” unless the contentclearly dictates otherwise.

Where multiple substituents are indicated as being attached to astructure, those substituents are independently selected. For example“ring A is optionally substituted with 1, 2 or 3 R_(q) groups” indicatesthat ring A is substituted with 1, 2 or 3 R_(q) groups, wherein theR_(q) groups are independently selected (i.e., can be the same ordifferent).

Compounds were named using Autonom 2000 4.01.305, which is availablefrom Beilstein Information Systems, Inc, Englewood, Colo.; ChemDrawv.10.0, (available from Cambridgesoft at 100 Cambridge Park Drive,Cambridge, Mass. 02140), or ACD Name pro, which is available fromAdvanced Chemistry Development, Inc., at 110 Yonge Street, 14^(th)floor, Toronto, Ontario, Canada M5c 1T4. Alternatively, the names weregenerated based on the IUPAC rules or were derived from names originallygenerated using the aforementioned nomenclature programs.

Where substituent groups are specified by their conventional chemicalformulae, written from left to right, they equally encompass thechemically identical substituents, which would result from writing thestructure from right to left. For example, “—CH₂O—” is intended to alsorecite “—OCH₂—”.

The term “alkyl,” by itself or as part of another substituent, means,unless otherwise stated, a straight or branched chain hydrocarbonradical, which can be fully saturated, mono- or polyunsaturated and caninclude di- and multivalent radicals, having the number of carbon atomsdesignated (i.e. C₁-C₁₀ means one to ten carbon atoms). Examples ofsaturated hydrocarbon radicals include, but are not limited to, groupssuch as methyl, ethyl, n-propyl, iso-propyl, n-butyl, tert-butyl,iso-butyl, sec-butyl, homologs and isomers of, for example, n-pentyl,n-hexyl, n-heptyl, n-octyl, and the like. An unsaturated alkyl group isone having one or more double bonds or triple bonds. Examples ofunsaturated alkyl groups include, but are not limited to, vinyl,2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl,3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butyryl, and thehigher homologs and isomers. Typically, an alkyl group will have from 1to 24 carbon atoms, with those groups having from 1 to 10 carbon atoms,from 1 to 8 carbon atoms or from 1 to 4 carbon atoms being preferred. A“lower alkyl” group is an alkyl group having from 1 to 4 carbon atoms.The term “alkyl” includes “alkylene” wherever appropriate, e.g., whenthe formula indicates that the alkyl group is divalent or whensubstituents are joined to form a ring.

The term “alkylene” by itself or as part of another substituent means adivalent radical derived from an alkane, as exemplified, but notlimited, by —CH₂CH₂CH₂CH₂—. Typically, an alkyl (or alkylene) group willhave from 1 to 24 carbon atoms, with those groups having 10 or fewercarbon atoms (e.g., 1 to 8 carbon atoms) being preferred in the presentinvention. A “lower alkylene” is an alkylene group, generally havingfrom 1 to 4 carbon atoms.

The term “alkenyl” by itself or as part of another substituent is usedin its conventional sense, and refers to a radical derived from analkene, as exemplified, but not limited, by substituted or unsubstitutedvinyl and substituted or unsubstituted propenyl. Typically, an alkenylgroup will have from 1 to 24 carbon atoms, with those groups having from1 to 10 carbon atoms being preferred.

The term “alkynyl” by itself or as part of another substituent is usedin its conventional sense, and refers to a radical derived from analkyne, as exemplified, but not limited, by substituted or unsubstitutedprop-1-ynyl, prop-2-ynyl (i.e., propargyl), and substituted orunsubstituted ethynyl. Typically, an alkynyl group will have from 1 to24 carbon atoms, with those groups having from 1 to 10 carbon atomsbeing preferred.

The terms “alkoxy,” “alkylamino” and “alkylthio” (or thioalkoxy) areused in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively.

The term “heteroalkyl,” by itself or in combination with another term,means, unless otherwise stated, a stable straight or branched chain, orcyclic hydrocarbon radical, or combinations thereof, consisting of thestated number of carbon atoms and at least one heteroatom selected fromthe group consisting of O, N, Si, S, B and P and wherein the nitrogenand sulfur atoms can optionally be oxidized and the nitrogen atom canoptionally be quaternized. The heteroatom(s) can be placed at anyinterior position of the heteroalkyl group or at the position at whichthe alkyl group is attached to the remainder of the molecule. Examplesof heteroalkyl groups include, but are not limited to, —CH₂—CH₂—O—CH₃,—CH₂—CH₂—NH—CH₃, —CH₂—CH₂—N(CH₃)—CH₃, —CH₂—S—CH₂—CH₃, —CH₂—CH₂,—S(O)—CH₃, —CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃, —Si(CH₃)₃, —CH₂—CH═N—OCH₃,and —CH═CH—N(CH₃)—CH₃. Up to two heteroatoms can be consecutive, suchas, for example, —CH₂—NH—OCH₃ and —CH₂—O—Si(CH₃)₃. Similarly, the term“heteroalkylene” by itself or as part of another substituent means adivalent radical derived from heteroalkyl, as exemplified, but notlimited by, —CH₂—CH₂—S—CH₂—CH₂— and —CH₂—S—CH₂—CH₂—NH—CH₂—. Forheteroalkylene groups, heteroatoms can also occupy either or both of thechain termini (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino,alkylenediamino, and the like). Still further, for alkylene andheteroalkylene linking groups, no orientation of the linking group isimplied by the direction in which the formula of the linking group iswritten. For example, the formula —CO₂R′ represents both —C(O)OR′ and—OC(O)R′. Typically, a heteroalkyl group will have from 2 to 24 atoms(2- to 24-membered), with those groups having from 2 to 10 atoms or from2 to 8 atoms being preferred. The term “heteroalkyl” includes“heteroalkylene” wherever appropriate, e.g., when the formula indicatesthat the heteroalkyl group is divalent or when substituents are joinedto form a ring.

The terms “cycloalkyl” and “heterocycloalkyl”, by themselves or incombination with other terms, represent, unless otherwise stated, cyclicversions of “alkyl” and “heteroalkyl”, respectively. Additionally, forheterocycloalkyl, a heteroatom can occupy the position at which theheterocycle is attached to the remainder of the molecule. A “cycloalkyl”or “heterocycloalkyl” substituent can be attached to the remainder ofthe molecule directly or through a linker. An exemplary linker isalkylene. Examples of cycloalkyl include, but are not limited to,cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl,and the like. The term “cycloalkyl” also includes bridged, polycyclic(e.g., bicyclic) structures, such as norbornane and adamantane.Typically, a cycloalkyl group will have from 3 to 24 carbon atoms, withthose groups having from 3 to 10 carbon atoms (e.g., C₃-C₈ cycloalkyl)being preferred.

In one example, a “heterocycloalkyl” group (also referred to as“heterocyclic group”, “heterocycle”, or “heterocyclyl”) is a carbocyclicring (e.g., 3- to 8-membered ring) containing at least one and up to 5heteroatoms (e.g., from 1 to 4 heteroatoms selected from nitrogen,oxygen and sulfur), or a fused ring system of 4- to 8-membered rings,containing at least one and up to 10 heteroatoms (e.g., from 1 to 5heteroatoms selected from nitrogen, oxygen and sulfur) in stablecombinations known to those of skill in the art. Attachment to theremainder of the molecule can be through either a carbon atom or aheteroatom. Exemplary heterocycloalkyl or heterocyclic groups of thepresent invention include morpholinyl, thiomorpholinyl, thiomorpholinylS-oxide, thiomorpholinyl S,S-dioxide, piperazinyl, homopiperazinyl,pyrrolidinyl, pyrrolinyl, imidazolidinyl, tetrahydropyranyl,piperidinyl, tetrahydrofuranyl, tetrahydrothienyl, piperidinyl,homopiperidinyl, homomorpholinyl, homothiomorpholinyl,homothiomorpholinyl S,S-dioxide, oxazolidinonyl, dihydropyrazolyl,dihydropyrrolyl, dihydropyrazolyl, dihydropyridyl, dihydropyrimidinyl,dihydrofuryl, dihydropyranyl, tetrahydrothienyl S-oxide,tetrahydrothienyl S,S-dioxide and homothiomorpholinyl S-oxide. Otherexamples of “heterocycloalkyl” include but are not limited to,1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl,3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl,tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl,1-piperazinyl, 2-piperazinyl, and the like.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl,” aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“halo(C₁-C₄)alkyl” is mean to include, but not be limited to,trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, andthe like.

The term “aryl” means, unless otherwise stated, a polyunsaturated,aromatic moiety that can be a single ring or a combination of multiplerings (e.g., from 1 to 3 rings), which are fused together or linkedcovalently and wherein at least one ring is aromatic. Non-limitingexamples of aryl groups include phenyl, 1-naphthyl, 2-naphthyl,4-biphenyl.

The term “heteroaryl” refers to aromatic moieties (e.g., a single ringor combination of multiple rings, fused or linked covalently) thatcontain from one to ten (preferably 1 to 5) heteroatoms selected from N,O, S, Si and B (preferably N, O and S), wherein the nitrogen and sulfuratoms are optionally oxidized, and the nitrogen atom(s) are optionallyquaternized. A heteroaryl group can be attached to the remainder of themolecule through a carbon- or heteroatom. Non-limiting examples ofheteroaryl groups include pyridyl, pyrimidinyl, quinolinyl,benzothienyl, indolyl, indolinyl, pyridazinyl, pyrazinyl, isoindolyl,isoquinolyl, quinazolinyl, quinoxalinyl, phthalazinyl, imidazolyl,isoxazolyl, pyrazolyl, oxazolyl, thiazolyl, indolizinyl, indazolyl,benzothiazolyl, benzimidazolyl, benzofuranyl, furanyl, thienyl,pyrrolyl, oxadiazolyl, thiadiazolyl, triazolyl, tetrazolyl,isothiazolyl, naphthyridinyl, isochromanyl, chromanyl,tetrahydroisoquinolinyl, isoindolinyl, isobenzotetrahydrofuranyl,isobenzotetrahydrothienyl, isobenzothienyl, benzoxazolyl, pyridopyridyl,benzotetrahydrofuranyl, benzotetrahydrothienyl, purinyl, benzodioxolyl,triazinyl, pteridinyl, benzothiazolyl, imidazopyridyl, imidazothiazolyl,dihydrobenzisoxazinyl, benzisoxazinyl, benzoxazinyl,dihydrobenzisothiazinyl, benzopyranyl, benzothiopyranyl, chromonyl,chromanonyl, pyridyl-N-oxide, tetrahydroquinolinyl, dihydroquinolinyl,dihydroquinolinonyl, dihydroisoquinolinonyl, dihydrocoumarinyl,dihydroisocoumarinyl, isoindolinonyl, benzodioxanyl, benzoxazolinonyl,pyrrolyl N-oxide, pyrimidinyl N-oxide, pyridazinyl N-oxide, pyrazinylN-oxide, quinolinyl N-oxide, indolyl N-oxide, indolinyl N-oxide,isoquinolyl N-oxide, quinazolinyl N-oxide, quinoxalinyl N-oxide,phthalazinyl N-oxide, imidazolyl N-oxide, isoxazolyl N-oxide, oxazolylN-oxide, thiazolyl N-oxide, indolizinyl N-oxide, indazolyl N-oxide,benzothiazolyl N-oxide, benzimidazolyl N-oxide, pyrrolyl N-oxide,oxadiazolyl N-oxide, thiadiazolyl N-oxide, triazolyl N-oxide, tetrazolylN-oxide, benzothiopyranyl S-oxide, benzothiopyranyl S,S-dioxide.Preferred heteroaryl groups include imidazolyl, pyrazolyl, thiadiazolyl,triazolyl, isoxazolyl, isothiazolyl, imidazolyl, thiazolyl, oxadiazolyl,and pyridyl. Other exemplary heteroaryl groups include 1-pyrrolyl,2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl,pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl,3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl,5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl,3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl,purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl,2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituentsfor each of the above noted aryl and heteroaryl ring systems areselected from the group of acceptable aryl group substituents describedbelow.

For brevity, the term “aryl” when used in combination with other terms(e.g., aryloxy, arylthioxy, arylalkyl) includes both aryl and heteroarylrings as defined above. Thus, the term “arylalkyl” is meant to includethose radicals in which an aryl group or heteroaryl group is attached toan alkyl group (e.g., benzyl, phenethyl, pyridylmethyl and the like).

Each of the above terms (e.g., “alkyl”, “cycloalkyl”, “heteroalkyl”,heterocycloalkyl”, “aryl” and “heteroaryl”) are meant to include bothsubstituted and unsubstituted forms of the indicated radical. Preferredsubstituents for each type of radical are provided below.

Substituents for the alkyl, cycloalkyl, heteroalkyl and heterocycloalkylradicals (including those groups often referred to as alkylene, alkenyl,heteroalkylene, heteroalkenyl, alkynyl, cycloalkenyl, andheterocycloalkenyl) are generically referred to as “alkyl groupsubstituents,” and they can be one or more of a variety of groupsselected from, but not limited to, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted aryl, substituted orunsubstituted heteroaryl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, —OR′, —SR′, ═O, ═NR′,═N—OR′, —NR′R″, -halogen, —SiR′R″R″′, —OC(O)R′, —C(O)R′, —CO₂R′,—CONR′R″, —OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R″′, —NR″C(O)₂R′,—NR—C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″,—NRSO₂R′, —N(SO₂R′)(SO₂R′), —CN and —NO₂ in a number ranging from zeroto (2 m′+1), where m′ is the total number of carbon atoms in suchradical. R′, R″, R″′ and R″″ each independently refer to hydrogen,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedaryl, e.g., aryl substituted with 1-3 halogens, substituted orunsubstituted alkyl, alkoxy or thioalkoxy groups, or arylalkyl groups.When a compound of the invention includes more than one R group, forexample, each of the R groups is independently selected as are each R′,R″, R′ and R′ groups when more than one of these groups is present. WhenR′ and R″ are attached to the same nitrogen atom, they can be combinedwith the nitrogen atom to form a 5-, 6-, or 7-membered ring. Forexample, —NR′R″ is meant to include 1-pyrrolidinyl and 4-morpholinyl.

Substituents for the aryl and heteroaryl groups are generically referredto as “aryl group substituents.” The substituents are selected from, forexample, substituted or unsubstituted alkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, substituted or unsubstituted heteroaryl, —OR′, ═O,═NR′, ═N—OR′, —NR′R″, —SR′, -halogen, —SiR′R″R″′, —OC(O)R′, —C(O)R′,—CO₂R′, —CONR′R″, —OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R″′, —NR″C(O)₂R′,—NR—C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″,—NRSO₂R′, —CN and —NO₂, —R′, —N₃, —CH(Ph)₂, fluoro(C₁-C₄)alkoxy, andfluoro(C₁-C₄)alkyl, in a number ranging from zero to the total number ofopen valences on the aromatic ring system, wherein R′, R″, R″′ and R′are independently selected from hydrogen, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted aryl and substituted or unsubstituted heteroaryl. When acompound of the invention includes more than one R group, for example,each of the R groups is independently selected as are each R′, R″, R′″and R″″ when more than one of these groups is present.

Two hydrogen atoms on adjacent atoms of the aryl or heteroaryl ring canoptionally be replaced with a substituent of the formula-T-C(O)—(CRR′)_(q)—U—, wherein T and U are independently —NR—, —O—,—CRR′— or a single bond, and q is an integer of from 0 to 3.Alternatively, two of the hydrogen atoms on adjacent atoms of the arylor heteroaryl ring can optionally be replaced with a substituent of theformula -A-(CH₂)_(r)—B—, wherein A and B are independently —CRR′—, —O—,—NR—, —S—, —S(O)—, —S(O)₂—, —S(O)₂NR′— or a single bond, and r is aninteger of from 1 to 4. One of the single bonds of the ring so formedcan optionally be replaced with a double bond. Alternatively, two of thehydrogen atoms on adjacent atoms of the aryl or heteroaryl ring canoptionally be replaced with a substituent of the formula—(CRR′)_(s)—X—(CR″R′″)_(d)—, where s and d are independently integers offrom 0 to 3, and X is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or —S(O)₂NR′—,wherein the substituents R, R′, R″ and R′″ are independently selectedfrom hydrogen and substituted or unsubstituted (C₁-C₆)alkyl.

As used herein, the term “acyl” describes the group —C(O)R. Exemplaryspecies for R include substituted or unsubstituted alkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted aryl, substitutedor unsubstituted heteroaryl, and substituted or unsubstitutedheterocycloalkyl.

As used herein, the term “fused ring system” means at least two rings,wherein each ring has at least 2 atoms in common with another ring.“Fused ring systems can include aromatic as well as non aromatic rings.Examples of “fused ring systems” are naphthalenes, indoles, quinolines,chromenes and the like.

As used herein, the term “heteroatom” includes oxygen (O), nitrogen (N),sulfur (S), silicon (Si), boron (B) and phosphorus (P). Preferredheteroatoms are O, S and N.

The symbol “R” is a general abbreviation that represents a substituentgroup. Exemplary substituent groups include substituted or unsubstitutedalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted aryl,substituted or unsubstituted heteroaryl, and substituted orunsubstituted heterocycloalkyl groups.

As used herein, the term “aromatic ring” or “non-aromatic ring” isconsistent with the definition commonly used in the art. For example,aromatic rings include phenyl and pyridyl. Non-aromatic rings includecyclohexanes.

The phrase “therapeutically effective amount” as used herein means thatamount of a compound, material, or composition of the present invention,which is effective for producing a desired therapeutic effect, at areasonable benefit/risk ratio applicable to any medical treatment. Forexample, a “therapeutically effective amount” can be an amount effectiveto reduce or lessen at least one symptom of the disease or conditionbeing treated or to reduce or delay onset of one or more clinicalmarkers or symptoms associated with the disease or condition, or tomodify or reverse the disease process.

The terms “treatment” or “treating” when referring to a disease orcondition, includes producing a therapeutic effect. Exemplarytherapeutic effects include delaying onset or reducing at least onesymptom associated with the disease, positively affecting (e.g.,reducing or delaying onset) of a clinical marker associated with thedisease and slowing or reversing disease progression.

The term “pharmaceutically acceptable salts” includes salts of thecompounds of the invention, which are prepared with nontoxic acids orbases, depending on the particular substituents found on the compoundsdescribed herein. When compounds of the present invention containrelatively acidic functionalities, base addition salts can be obtainedby contacting the neutral form of such compounds with a sufficientamount of the desired base, either neat or in a suitable inert solvent.Examples of pharmaceutically acceptable base addition salts includesodium, potassium, calcium, ammonium, organic amino, magnesium salts andthe like. When compounds of the present invention contain relativelybasic functionalities (e.g., amines), acid addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,diphosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic and the like, as well as the saltsderived from relatively nontoxic organic acids like formic, acetic,propionic, isobutyric, malic, maleic, malonic, benzoic, succinic,suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic,p-tolylsulfonic, citric, tartaric, methanesulfonic,2-hydroxyethylsulfonic, salicylic, stearic and the like. Also includedare salts of amino acids such as arginate and the like, and salts oforganic acids like glucuronic or galactunoric acids and the like (see,for example, Berge et al., Journal of Pharmaceutical Science, 1977, 66:1-19). Similarly, pharmaceutically acceptable cations include, but arenot limited to sodium, potassium, calcium, aluminum, lithium andammonium. Certain specific compounds of the present invention containboth basic and acidic functionalities that allow the compounds to beconverted into either base or acid addition salts.

When a residue is defined as “O⁻”, e.g., in “—COO⁻”, then the formula ismeant to optionally include an organic or inorganic cationic counterion.In one example, the resulting salt form of the compound ispharmaceutically acceptable. Further, when a compound of the inventionincludes an acidic group, such as a carboxylic acid group, e.g., writtenas the substituent “—COOH”, “—CO₂H” or “—C(O)₂H”, then the formula ismeant to optionally include the corresponding “de-protonated” form ofthat acidic group, e.g., “—COO⁻”, “—CO₂ ⁻” or “—C(O)₂ ⁻”, respectively.

The neutral forms of the compounds can be regenerated, for example, bycontacting the salt with a base or acid and isolating the parentcompound in the conventional manner. The parent form of the compound candiffer from the various salt forms in certain physical properties, suchas solubility in polar solvents, but otherwise the salts are equivalentto the parent form of the compound for the purposes of the presentinvention.

In addition to salt forms, the present invention provides compounds,which are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Non-limiting examples of “pharmaceutically acceptablederivative” or “prodrug” include pharmaceutically acceptable esters,phosphate esters, sulfonate esters or salts thereof as well as otherderivatives of a compound of this invention which, upon administrationto a recipient, is capable of providing, either directly or indirectly,a compound of this invention. Particularly favored derivatives orprodrugs are those that increase the bioavailability of the compounds ofthis invention when such compounds are administered to a mammal (e.g.,by allowing an orally administered compound to be more readily absorbedinto the blood) or which enhance delivery of the parent compound to abiological compartment (e.g., the brain or lymphatic system) relative tothe parent species.

Prodrugs include a variety of esters (i.e., carboxylic acid ester).Ester groups, which are suitable as prodrug groups are generally knownin the art and include benzyloxy, di(C₁-C₆)alkylaminoethyloxy,acetoxymethyl, pivaloyloxymethyl, phthalidoyl, ethoxycarbonyloxyethyl,5-methyl-2-oxo-1,3-dioxol-4-yl methyl, and (C₁-C₆)alkoxy esters,optionally substituted by N-morpholino and amide-forming groups such asdi(C₁-C₆)alkylamino. Preferred ester prodrug groups include C₁-C₆ alkoxyesters. Those skilled in the art will recognize various syntheticmethodologies that may be employed to form pharmaceutically acceptableprodrugs of the compounds of the invention (e.g., via esterification ofa carboxylic acid group).

In an exemplary embodiment, the prodrug is suitable fortreatment/prevention of those diseases and conditions that require thedrug molecule to cross the blood brain barrier. In a preferredembodiment, the prodrug enters the brain, where it is converted into theactive form of the drug molecule. In another example, a prodrug is usedto enable an active drug molecule to reach the inside of the eye aftertopical application of the prodrug to the eye. Additionally, prodrugscan be converted to the compounds of the present invention by chemicalor biochemical methods in an ex vivo environment. For example, prodrugscan be slowly converted to the compounds of the present invention whenplaced in a transdermal patch reservoir with a suitable enzyme orchemical reagent.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are encompassedwithin the scope of the present invention. Certain compounds of thepresent invention can exist in multiple crystalline or amorphous forms(“polymorphs”). In general, all physical forms are of use in the methodscontemplated by the present invention and are intended to be within thescope of the present invention. “Compound or a pharmaceuticallyacceptable salt, hydrate, polymorph or solvate of a compound” intendsthe inclusive meaning of “or”, in that materials meeting more than oneof the stated criteria are included, e.g., a material that is both asalt and a solvate is encompassed.

The compounds of the present invention can contain unnatural proportionsof atomic isotopes at one or more of the atoms that constitute suchcompounds. For example, the compounds can be radiolabeled withradioactive isotopes, such as for example tritium (³H), iodine-125(¹²⁵I) or carbon-14 (¹⁴C). All isotopic variations of the compounds ofthe present invention, whether radioactive or not, are intended to beencompassed within the scope of the present invention.

The term “solvate” is intended to refer to a complex formed bycombination of solute molecules or ions with solvent molecules. Thesolvent can be an organic compound, an inorganic compound, or a mixtureof both. Exemplary solvents for the formation of solvates include, butare not limited to, methanol, N,N-dimethylformamide, tetrahydrofuran,dimethylsulfoxide, toluene, and water. In one embodiment, solventshaving a higher boiling point, such as for example, DMF, DMA, and thelike.

Compositions Including Stereoisomers

Compounds of the invention can exist in particular geometric orstereoisomeric forms. The invention contemplates all such compounds,including cis- and trans-isomers, (−)- and (+)-enantiomers,diastereomers, (D)-isomers, (L)-isomers, the racemic mixtures thereof,and other mixtures thereof, such as enantiomerically ordiastereomerically enriched mixtures, as falling within the scope of theinvention. Additional asymmetric carbon atoms can be present in asubstituent such as an alkyl group. All such isomers, as well asmixtures thereof, are intended to be included in this invention. Whenthe compounds described herein contain olefinic double bonds or othercenters of geometric asymmetry, and unless specified otherwise, it isintended that the compounds include both E and Z geometric isomers.Likewise, all tautomeric forms and mixtures of tautomers are included.

Optically active (R)- and (S)-isomers and d and l isomers can beprepared using chiral synthons or chiral reagents, or resolved usingconventional techniques. Resolution of the racemates can beaccomplished, for example, by conventional methods such ascrystallization in the presence of a resolving agent; chromatography,using, for example a chiral HPLC column; or derivatizing the racemicmixture with a resolving reagent to generate diastereomers, separatingthe diastereomers via chromatography, and removing the resolving agentto generate the original compound in enantiomerically enriched form. Anyof the above procedures can be repeated to increase the enantiomericpurity of a compound. If, for instance, a particular enantiomer of acompound of the present invention is desired, it can be prepared byasymmetric synthesis, or by derivatization with a chiral auxiliary,where the resulting diastereomeric mixture is separated and theauxiliary group cleaved to provide the pure desired enantiomers.Alternatively, where the molecule contains a basic functional group,such as an amino group, or an acidic functional group, such as acarboxyl group, diastereomeric salts can be formed with an appropriateoptically active acid or base, followed by resolution of thediastereomers thus formed by fractional crystallization orchromatographic means known in the art, and subsequent recovery of thepure enantiomers. In addition, separation of enantiomers anddiastereomers is frequently accomplished using chromatography employingchiral, stationary phases, optionally in combination with chemicalderivatization (e.g., formation of carbamates from amines).

As used herein, the term “chiral”, “enantiomerically enriched” or“diastereomerically enriched” refers to a compound having anenantiomeric excess (ee) or a diastereomeric excess (de) of greater thanabout 50%, preferably greater than about 70% and more preferably greaterthan about 90%. In general, higher than about 90% enantiomeric ordiastereomeric excess is particularly preferred, e.g., thosecompositions with greater than about 95%, greater than about 97% andgreater than about 99% ee or de.

The terms “enantiomeric excess” and “diastereomeric excess” are usedinterchangeably herein. Compounds with a single stereocenter arereferred to as being present in “enantiomeric excess”, those with atleast two stereocenters are referred to as being present in“diastereomeric excess”.

The term “enantiomeric excess” is related to the older term “opticalpurity” in that both are measures of the same phenomenon. The value ofee will be a number from 0 to 100, zero being racemic and 100 beingenantiomerically pure. A compound which in the past might have beencalled 98% optically pure is now more precisely characterized by 96% ee.A 90% ee reflects the presence of 95% of one enantiomer and 5% of theother(s) in the material in question.

Hence, in one embodiment, the invention provides a composition includinga first stereoisomer and at least one additional stereoisomer of acompound of the invention. The first stereoisomer can be present in adiastereomeric or enantiomeric excess of at least about 80%, preferablyat least about 90% and more preferably at least about 95%. In aparticularly preferred embodiment, the first stereoisomer is present ina diastereomeric or enantiomeric excess of at least about 96%, at leastabout 97%, at least about 98%, at least about 99% or at least about99.5%. In another embodiment, the compound of the invention isenantiomerically or diastereomerically pure (diastereomeric orenantiomeric excess is about 100%). Enantiomeric or diastereomericexcess can be determined relative to exactly one other stereoisomer, orcan be determined relative to the sum of at least two otherstereoisomers. In an exemplary embodiment, enantiomeric ordiastereomeric excess is determined relative to all other detectablestereoisomers, which are present in the mixture. Stereoisomers aredetectable if a concentration of such stereoisomer in the analyzedmixture can be determined using common analytical methods, such aschiral HPLC.

Integrins are a large family of homologous transmembrane linker proteinsthat are the principal receptors on animal cells for binding mostextracellular matrix proteins, such as collagen, fibronectin, andlaminin. The integrins are heterodimers comprised of an α chain and a βchain. To date, twenty different integrin heterodimers, made from 9different α subunits and 14 different β subunits, have been identified.The term “α 4 integrins” refers to the class of heterodimer,enzyme-linked cell-surface receptors that contain the α 4 subunit pairedwith any of the β subunits. VLA-4 is an example of an α 4 integrin, andis a heterodimer of the α 4 and β1 subunits, and is also referred to asα 4 β1 integrin.

Compositions

In various aspects, the invention provides a compound having a structureaccording to Formula (Ia), Formula (Ia′), Formula (Ib) or Formula (Ic):

or a salt or solvate thereof, wherein m is an integer selected from 0 to4, n is an integer selected from 0 to 4, p is an integer selected from 0to 6 and m1 is an integer selected from 0 to 3. In one example, n isselected from 1 to 4. In another example, n is selected from 1 and 2. Inyet another example, n is 1. In a further example, m is 0 or 1. Inanother example, p is 0 or 1.

In Formula (Ia), Formula (Ia′), Formula (Ib) or Formula (Ic), ring A isa member selected from substituted or unsubstituted aryl (e.g., phenyl,naphthyl) and substituted or unsubstituted heteroaryl (e.g., pyridyl,thiophene, thiazole, imidazolyl).

In Formula (Ia), Formula (Ia′), Formula (Ib) or Formula (Ic), Cy is amember selected from substituted or unsubstituted cycloalkyl (e.g.,C₃-C₁₀ cycloalkyl), substituted or unsubstituted heterocycloalkyl (e.g.,3- to 10-membered heterocycloalkyl), substituted or unsubstituted aryl(e.g., phenyl or naphthyl) and substituted or unsubstituted heteroaryl(e.g., pyridyl). In one example in the above structures, Cy issubstituted or unsubstituted aryl (e.g., phenyl or naphthyl) orsubstituted or unsubstituted heteroaryl (e.g., pyridyl). In a particularexample, Cy in the above structures is a member selected fromsubstituted or unsubstituted phenyl (e.g., substituted or unsubstituted4-chloro-phenyl or substituted or unsubstituted 4-phenoxyphenyl),substituted or unsubstituted naphthyl, substituted or unsubstitutedbiphenyl (e.g., substituted or unsubstituted biphenyl-4-yl) andsubstituted or unsubstituted pyridyl (e.g., substituted or unsubstituted4-pyridyl).

In Formula (Ia), Formula (Ia′), Formula (Ib) or Formula (Ic), each R¹,each R², R^(1a) and R^(1b) are members independently selected from H,substituted or unsubstituted alkyl (e.g., C₁-C₁₀ alkyl), substituted orunsubstituted heteroalkyl (e.g., 2- to 10-membered heteroalkyl),substituted or unsubstituted cycloalkyl (e.g., C₃-C₁₀ cycloalkyl),substituted or unsubstituted heterocycloalkyl (e.g., 3- to 10-memberedheterocycloalkyl), substituted or unsubstituted aryl (e.g., substitutedor unsubstituted phenyl or naphthyl), substituted or unsubstitutedheteroaryl (e.g., substituted or unsubstituted pyridyl), nitro, CN,halogen (e.g., I, F, Cl or Br), OR¹², SR¹², NR¹²R¹³, C(O)R¹⁴,C(O)NR¹²R¹³, OC(O)NR¹²R¹³, C(O)OR¹², NR¹⁵C(O)R¹⁴, NR¹⁵C(O)OR¹²,NR¹⁵C(O)NR¹²R¹³, NR¹⁵C(S)NR¹²R¹³, NR¹⁵S(O)₂R¹⁴, S(O)₂NR¹²R¹³ andS(O)_(z)R¹⁴, wherein z is 1 or 2. R¹², R¹³ and R¹⁵ are membersindependently selected from H, acyl, substituted or unsubstituted alkyl(e.g., C₁-C₁₀ alkyl), substituted or unsubstituted heteroalkyl (e.g., 2-to 10-membered heteroalkyl), substituted or unsubstituted aryl,substituted or unsubstituted heteroaryl, substituted or unsubstitutedcycloalkyl (e.g., C₃-C₁₀ cycloalkyl) and substituted or unsubstitutedheterocycloalkyl (e.g., 3- to 10-membered heterocycloalkyl). R¹⁴ is amember independently selected from substituted or unsubstituted alkyl(e.g., C₁-C₁₀ alkyl), substituted or unsubstituted heteroalkyl (e.g., 2-to 10-membered heteroalkyl), substituted or unsubstituted aryl,substituted or unsubstituted heteroaryl, substituted or unsubstitutedcycloalkyl (e.g., C₃-C₁₀ cycloalkyl) and substituted or unsubstitutedheterocycloalkyl (e.g., 3- to 10-membered heterocycloalkyl). R¹² andR¹³, together with the nitrogen atoms to which they are attached, areoptionally joined to form a 4- to 7-membered ring. Two adjacent R¹,together with the atoms to which they are attached, are optionallyjoined to form a 5- to 7-membered ring. Two adjacent R², together withthe atoms to which they are attached, are optionally joined to form a 5-to 7-membered ring.

In Formula (Ia), Formula (Ia′), Formula (Ib) or Formula (Ic), R³, R⁴ andR⁵ are members independently selected from H, substituted orunsubstituted alkyl (e.g., C₁-C₄ alkyl), halogen (e.g., I, F, Cl, Br),substituted or unsubstituted aryl, —O-aryl, and CN. In a particularexample in the above structures, R³, R⁴ and R⁵ are each H.

In Formula (Ia), Formula (Ia′), Formula (Ib) or Formula (Ic), R⁶ is amember selected from H, substituted or unsubstituted alkyl (e.g., C₁-C₆alkyl), substituted or unsubstituted heteroalkyl (e.g., 2- to 6-memberedheteroalkyl), substituted or unsubstituted cycloalkyl (e.g., C₃-C₈cycloalkyl), substituted or unsubstituted heterocycloalkyl (e.g., 3- to8-membered heterocycloalkyl), substituted or unsubstituted aryl (e.g.,substituted or unsubstituted phenyl), substituted or unsubstitutedheteroaryl (e.g., substituted or unsubstituted pyridyl), NR¹²R¹³,C(O)R¹⁴, C(O)NR¹²R¹³, C(O)OR¹² and S(O)_(z)R¹⁴, wherein z is 1 or 2 andR¹², R¹³ and R¹⁴ are defined as herein above. In one example, R⁶ is amember selected from H and substituted or unsubstituted alkyl (e.g.,C₁-C₄ alkyl). In a particular example in the above structures, R⁶ is H.

In Formula (Ic), E is selected from O, S, C(O), S(O)₂ and NR⁴⁰, whereinR⁴⁰ is a member selected from substituted or unsubstituted alkyl (e.g.,C₁-C₁₀ alkyl). In one example, R⁴⁰ is unsubstituted C₁-C₄ alkyl (e.g.,methyl or ethyl).

In one example, in Formula (Ia) or (Ia′), n is 1 and R³, R⁴, R⁵ and R⁶are each H. In another example, in Formula (Ib), R³, R⁴, R⁵ and R⁶ areeach H. Hence, an exemplary compound of the invention has a structureaccording to Formula (IIa), Formula (IIa′), or Formula (IIb):

wherein m, p, Cy, ring A, R¹, R^(1a), R² and R⁶ are defined as forFormula (Ia), Formula (Ia′), and Formula (Ib), above. In one example, inFormula (IIa), Formula (IIa′) or Formula (IIb), ring A is substituted orunsubstituted phenyl or substituted or unsubstituted thiophene. Inanother example, in Formula (IIa), Formula (IIa′) or Formula (IIb), Cyis a member selected from substituted or unsubstituted aryl (e.g.,phenyl or naphthyl) and substituted or unsubstituted heteroaryl (e.g.,pyridyl).

Ring A

In one example, in Formula (Ia), Formula (Ia′), Formula (Ib), Formula(IIa), Formula (IIa′), or Formula (IIb), ring A is a 5- or 6-memberedaromatic or heteroaromatic ring. Exemplary rings for A include phenyl,pyridine, thiophene, thiazole and oxazole. In another example, A isphenyl or thiophene and the compound of the invention has a structureaccording to Formula (IIIa), Formula (IIIa′), or Formula (IIIb):

or a salt or solvate thereof, wherein m, Cy, R¹, R², R³, R⁴, R⁵ and R⁶are defined as above. The integer q is selected from 0 to 4. The integerr is selected from 0 to 2. In one example, in the above formulae, eachof R³, R⁴, R⁵ and R⁶ is H.

In Formula (IIIa) or (IIIa′), X¹, X², X³ and X⁴ are membersindependently selected from N and CR², wherein each R² is independentlydefined as above.

In Formula (IIIb), Y¹, Y² and Y³ are members independently selected fromS, O, N, NR^(2a) and CR², with the proviso that at least one of Y¹, Y²and Y³ is other than CR². Each R² is defined as hereinabove. R^(2a) is amember selected from H, substituted or unsubstituted alkyl (e.g.,substituted or unsubstituted C₁-C₈ alkyl), substituted or unsubstitutedheteroalkyl (e.g., substituted or unsubstituted 2- to 8-memberedheteroalkyl), substituted or unsubstituted cycloalkyl (e.g., substitutedor unsubstituted C₃-C₈ cycloalkyl), substituted or unsubstitutedheterocycloalkyl (e.g., substituted or unsubstituted 3- to 8-memberedheterocycloalkyl), substituted or unsubstituted aryl, and substituted orunsubstituted heteroaryl. In one example, R^(2a) is selected from H andsubstituted or unsubstituted C₁-C₆ alkyl (e.g., methyl or ethyl).

In a further example, the compounds of the invention have a structureaccording to Formula (IV), Formula (V) or Formula (VI):

or a tautomer, salt or solvate thereof, wherein m, Cy, R¹, R², R³, R⁴,R⁵ and R⁶ are defined as above. The integer q is selected from 0 to 4.The integer r is selected from 0 to 2.

In one example according to any of the above embodiments, R⁶ is H.

Unexpectedly, the inventors have discovered that in vitro biologicalactivity is generally higher, when the pyridinone (pyridone) ring isunsubstituted or substituted with a small substituent, such as F. Hence,in another example according to any of the above embodiments, each ofR³, R⁴ and R⁵ is independently selected from H, halogen (e.g., F, Cl),CN and C₁-C₃ alkyl. Preferably, each of R³, R⁴ and R⁵ is independentlyselected from H and F. In a further example, each of R³, R⁴ and R⁵ is H.In yet another example, each of R³, R⁴, R⁵ and R⁶ is H and the compoundof the invention has a structure according to Formula (IVa), Formula(Va), or Formula (VIa):

or a tautomer, salt or solvate thereof, wherein m, Cy, R¹, R² aredefined as above. The integer q is selected from 0 to 4. The integer ris selected from 0 to 2.

In one example, the compound of the invention has a structure accordingto Formula (IVb) or Formula (IVc):

or a salt or solvate thereof, wherein Cy, R¹, R² and m are defined asherein above.

Ring Cy

Cy in any of the above formulae can be a ring or fused ring system. Inone embodiment, Cy in any of the above formulae is a member selectedfrom substituted or unsubstituted cycloalkyl (e.g., C₃-C₈ cycloalkyl),substituted or unsubstituted heterocycloalkyl (e.g., 3- to 8-memberedheterocycloalkyl), substituted or unsubstituted aryl and substituted orunsubstituted heteroaryl. In a particular example, Cy is a memberselected from substituted or unsubstituted aryl (e.g., phenyl, biphenyl,naphthyl) and substituted or unsubstituted heteroaryl (e.g., pyridyl,quinolinyl, quinazolinyl). In another example, Cy is selected from6-membered aryl and 6-membered heteroaryl moieties of the formula:

wherein X⁵, X⁶, X⁷, X⁸ and X⁹ are members independently selected from Nand CR¹⁶, wherein each R¹⁶ is independently selected from aryl groupsubstituents. In one example, each R¹⁶ is a member independentlyselected from H, substituted or unsubstituted alkyl (e.g., substitutedor unsubstituted C₁-C₈ alkyl), substituted or unsubstituted heteroalkyl(e.g., substituted or unsubstituted 2- to 8-membered heteroalkyl),substituted or unsubstituted cycloalkyl (e.g., substituted orunsubstituted C₃-C₈ cycloalkyl), substituted or unsubstitutedheterocycloalkyl (e.g., substituted or unsubstituted 3- to 8-memberedheterocycloalkyl), substituted or unsubstituted aryl, substituted orunsubstituted heteroaryl, halogen, CN, OR¹⁷, SR¹⁷, NR¹⁷R¹⁸, C(O)R¹⁹,C(O)NR¹⁷R¹⁸, OC(O)NR¹⁷R¹⁸, C(O)OR¹⁷, NR²⁰C(O)R¹⁹, NR²⁰C(O)OR¹⁷,NR²⁰C(O)NR¹⁷R¹⁸, NR²⁰C(S)NR¹⁷R¹⁸, NR²⁰S(O)₂R¹⁹, S(O)₂NR¹⁷R¹⁸ andS(O)_(p)R¹⁹, wherein p is 1 or 2.

Each R¹⁷, each R¹⁸ and each R²⁰ is a member independently selected fromH, acyl, substituted or unsubstituted alkyl (e.g., substituted orunsubstituted C₁-C₈ alkyl), substituted or unsubstituted heteroalkyl(e.g., substituted or unsubstituted 2- to 8-membered heteroalkyl),substituted or unsubstituted cycloalkyl (e.g., substituted orunsubstituted C₃-C₈ cycloalkyl), substituted or unsubstitutedheterocycloalkyl (e.g., substituted or unsubstituted 3- to 8-memberedheterocycloalkyl), substituted or unsubstituted aryl, and substituted orunsubstituted heteroaryl. R¹⁷ and R¹⁸, together with the nitrogen atomto which they are attached, are optionally joined to form a 5- to7-membered ring. Each R¹⁹ is a member independently selected fromsubstituted or unsubstituted alkyl (e.g., substituted or unsubstitutedC₁-C₈ alkyl), substituted or unsubstituted heteroalkyl (e.g.,substituted or unsubstituted 2- to 8-membered heteroalkyl), substitutedor unsubstituted cycloalkyl (e.g., substituted or unsubstituted C₃-C₈cycloalkyl), substituted or unsubstituted heterocycloalkyl (e.g.,substituted or unsubstituted 3- to 8-membered heterocycloalkyl),substituted or unsubstituted aryl, and substituted or unsubstitutedheteroaryl. Adjacent R¹⁶, together with the carbon atoms to which theyare attached, are optionally joined to form a 5- to 7-membered ring.

In one example, the compound of the invention has a structure accordingto Formula (VII):

wherein m, n, p, A, R¹, R², R³, R⁴, R⁵ and R⁶ are defined as above. X⁵,X⁶, X⁷, X⁸ and X⁹ are also defined as herein above.

In one example, the compound of the invention has a structure accordingto Formula (VIIIa) or Formula (VIIIb):

wherein m, p, R¹, R², R³, R⁴, R⁵ and R⁶ are defined as above. Y¹, Y²,Y³, the integer r, X¹, X², X³, X⁴, X⁵, X⁶, X⁷, X⁸ and X⁹ are defined asherein above.

In a further example, the compounds of the invention have a structureaccording to Formula (IX), Formula (X) or Formula (XI):

or a tautomer, salt or solvate thereof, wherein m, q, R¹, R², R³, R⁴, R⁵and R⁶ are defined as above. X⁵, X⁶, X⁷, X⁸ and X⁹ are defined as hereinabove.

In a further embodiment, Cy or the moiety:

in any of the above formulae is a member selected from:

wherein w is an integer selected from 0 to 5, s is an integer selectedfrom 0 to 4, t is an integer selected from 0 to 3 and u is an integerselected from 0 to 2.

In another example, Cy is a fused ring system, which includes at leastone of the above rings. In one example, Cy is a member selected frombenzo- or pyrido-imidazole, benzo- or pyrido-oxazole, benzo- orpyrido-thiazole, benzo- or pyrido-isoxazole and benzo- orpyrido-isothiazole.

In a further example, Cy in any of the above embodiments is4-substituted or 3-substituted phenyl or pyridyl. For example, Cy has astructure selected from:

wherein u is an integer selected from 0 to 4 and v is an integerselected from 0 to 3. R^(16a) is defined as R¹⁶ herein above with thedifference that R^(16a) is other than H. In one example, R^(16a) in theabove structures is OR¹⁷, wherein R¹⁷ is defined as herein above. In oneexample, R¹⁷ is a member selected from substituted or unsubstitutedalkyl. In a particular example, R^(16a) is a member selected frommethoxy and ethoxy. Each R^(20a) in the above structures isindependently selected and is defined as R¹⁶ herein above. In oneexample, each R^(20a) in the above structures is H.

Other exemplary rings for Cy include:

wherein R^(16a) and R^(20a) are defined as herein above, except thatboth, R^(16a) and R^(20a), are other than H. R^(16a) and R^(20a),together with the atoms to which they are attached, are optionallyjoined to form a 5- to 7-membered ring.

In a particular example, the compound of the invention has a structureaccording to Formula (XII):

or a salt or solvate thereof, wherein ring A, R¹, R², R³, R⁴, R⁵, R⁶, m,n, p, s, X⁷ and R¹⁶ are defined as herein above.

In another example, the compound of the invention has a structureaccording to Formula (XIIa) or Formula (XIIb):

wherein ring A, R¹, R², m, p, q, s, X⁷ and R¹⁶ are defined as hereinabove. In one example, X⁷ in the above structures is a member selectedfrom N and CR^(16a), wherein R^(16a) is defined as R¹⁶ herein above,except that R^(16a) is other than H.

In yet another example according to any of the above embodiments, Cy issubstituted or unsubstituted 4-pyridyl or 4-substituted phenyl.Exemplary compounds of the invention have a structure according toFormula (XIIc) or Formula (XIId):

wherein R¹, R², R^(16a), m and q are defined as herein above.

In one example according to any of the above embodiments, R^(16a) isselected from halogen (e.g., F, Cl, Br), halogen substituted lower C₁-C₄alkyl (e.g., CF₃), lower alkyl (e.g., methyl, ethyl), substituted orunsubstituted aryl (e.g., substituted or unsubstituted phenyl orsubstituted or unsubstituted pyridyl), substituted or unsubstitutedheteroaryl (e.g., furan, thiophene, thiazole), OR¹⁷, wherein R¹⁷ isdefined as herein above. In a particular example, R¹⁷ in OR¹⁷ isselected from CF₃, substituted or unsubstituted aryl (e.g., phenyl) andsubstituted or unsubstituted heteroaryl. In another example, R^(16a) isNR¹⁷R¹⁸, wherein R¹⁷ and R¹⁸ are defined as herein above. In oneexample, in NR¹⁷R¹⁸, R¹⁷ is H and R¹⁸ is selected from substituted orunsubstituted phenyl. In another example according to any of the aboveembodiments, R^(16a) is a member selected from halogen (e.g., F, Cl,Br), substituted or unsubstituted phenyl and substituted orunsubstituted phenyloxy.

In yet another example, Cy is a member selected from:

wherein v is an integer selected from 0 to 3 and x is an integerselected from 0 to 4. A is O, S or NR³⁰, wherein R³⁰ is a memberselected from H and C₁-C₄ alkyl (e.g., methyl). Each R^(20a) in theabove structures is independently selected and is defined as hereinabove. In one example, each R^(20a) in the above structures is H. X⁵,X⁶, X⁷, X⁸ and X⁹ are defined as herein above. In one example, X⁶, X⁷,X⁸ and X⁹ are independently selected from N and CH.

In yet another example, Cy is a member selected from:

wherein each v is an integer independently selected from 0 to 3, x is aninteger selected from 0 to 4 and y is an integer selected from 0 to 5.Y^(5a) is selected from O and S. R^(20a) and R³⁰ are defined as hereinabove. R^(20b) is defined as R^(20a), wherein R^(20a) and R^(20b) areindependently selected. In one example in the above structures eachR^(20a) is H. In another example, each R^(20b) in the above structuresis H. In yet another example, each R^(20a) and each R^(20b) in the abovestructures is H. In yet another example, v in (R^(20b))_(v) is 1 andR^(20b) is methoxy.

In another example, Cy is a 5-membered heteroaryl moiety of the formula:

wherein Y⁴, Y⁵, Y⁶ and Y⁷ are members independently selected from S, O,N, NR²² and CR²³, with the proviso that at least one of Y¹, Y² and Y³ isother than CR²³. Each R²² and each R²³ is a member independentlyselected from aryl group substituents. In one example, each R²² and eachR²³ is a member independently selected from H, substituted orunsubstituted alkyl (e.g., substituted or unsubstituted C₁-C₈ alkyl),substituted or unsubstituted heteroalkyl (e.g., substituted orunsubstituted 2- to 8-membered heteroalkyl), substituted orunsubstituted cycloalkyl (e.g., substituted or unsubstituted C₃-C₈cycloalkyl), substituted or unsubstituted heterocycloalkyl (e.g.,substituted or unsubstituted 3- to 8-membered heterocycloalkyl),substituted or unsubstituted aryl, and substituted or unsubstitutedheteroaryl. In one example, R²² and R²³ are members independentlyselected from H and substituted or unsubstituted C₁-C₆ alkyl (e.g.,methyl or ethyl). Each R²³ can further be selected from OR¹⁷, SR″,NR¹⁷R¹⁸, C(O)R¹⁹, C(O)NR¹⁷R¹⁸, OC(O)NR¹⁷R¹⁸, C(O)OR¹⁷, NR²⁰C(O)OR¹⁹,NR²⁰C(O)OR¹⁷, NR²⁰C(O)NR¹⁷R¹⁸, NR²⁰C(S)NR¹⁷R¹⁸, NR²⁰S(O)₂R¹⁹,S(O)₂NR¹⁷R¹⁸ and S(O)_(p)R¹⁹, wherein p is 1 or 2, wherein R¹⁷, R¹⁸, R¹⁹and R²⁰ are defined as herein above.

In one example, the compound of the invention has a structure accordingto Formula (XIII):

wherein m, n, p, A, R¹, R², R³, R⁴, R⁵ and R⁶ are defined above. Y⁴, Y⁵,Y⁶ and Y⁷ are also defined as herein above.

In one example, the compound of the invention has a structure accordingto Formula (XIIIa) or Formula (XIIIb):

wherein m, q, r, R¹, R², R³, R⁴, R⁵ and R⁶ are defined as above. X¹, X²,X³, X⁴, Y¹, Y², Y³, Y⁴, Y⁵, Y⁶ and Y⁷ are defined as herein above.

In a further example, the compounds of the invention have a structureaccording to Formula (XIV), Formula (XV) or Formula (XVI):

or a tautomer, salt or solvate thereof, wherein m, q, r, R¹, R², R³, R⁴,R⁵ and R⁶ are defined as above. Y¹, Y², Y³, Y⁴, Y⁵, Y⁶ and Y⁷ aredefined as herein above.

In one example, Cy or the moiety

in any of the above formulae and embodiments, is a member selected from:

or a tautomer or mixture of tautomers thereof, wherein R²² and R²³ aredefined as hereinabove.

In one embodiment, in the above structures, each R²³ is a memberindependently selected from H, C₁-C₄ alkyl and alkoxy.

Exemplary compounds of the invention and their in vitro biologicalactivities are listed in the table of FIG. 1.

In Vitro Activities

Certain compounds of the invention exhibit various in vitro biologicalactivities. Specifically, certain compounds of the invention inhibitbinding of alpha4 integrins to their respective ligands. Those compoundsare termed alpha4 antagonists. For example, compounds of the inventioninhibit binding of alpha4 integrins (e.g., alpha4beta7 or VLA-4) totheir natural ligands, such as fibronectin, VCAM-1 or MadCAM. In vitroassays for the determination of such activities are known in the art(see e.g., WO2000/51974, incorporated herein by reference). Exemplaryassay formats are described herein (see e.g., Examples 32-38).

In one example, the compounds of the invention inhibit binding ofalpha4beta7 to MadCAM in a MadCAM adhesion assay (e.g., at least one ofthe assays described in Examples 32 and 33) with an IC₅₀ of less thanabout 50 μM, less than about 40 μM, less than about 30 μM, less thanabout 20 μM or less than about 10 μM.

In another example, the compounds of the invention inhibit binding ofalpha4beta7 to MadCAM in a MadCAM adhesion assay with an IC₅₀ of lessthan about 9 μM, less than about 8 μM, less than about 7 μM, less thanabout 6 μM, less than about 5 μM, less than about 4 μM, less than about3 μM, less than about 2 μM, or less than about 1 μM. In yet anotherexample, the compounds of the invention inhibit binding of alpha4beta7to MadCAM in a MadCAM adhesion assay with an IC₅₀ of less than about 0.9μM, less than about 0.8 μM, less than about 0.7 μM, less than about 0.6μM, less than about 0.5 μM, less than about 0.4 μM, less than about 0.3μM, less than about 0.2 μM. In a particular example, the compounds ofthe invention inhibit binding of alpha4beta7 to MadCAM in a MadCAMadhesion assay with an IC₅₀ of less than about 0.1 μM (100 nM). Inanother particular example, the compounds of the invention inhibitbinding of alpha4beta7 to MadCAM in a MadCAM adhesion assay with an IC₅₀of less than about 90 nM, less than about 80 nM, less than about 70 nM,less than about 60 nM, less than about 50 nM, less than about 40 nM,less than about 30 nM or less than about 20 nM. In another particularexample, the compounds of the invention inhibit binding of alpha4beta7to MadCAM in a MadCAM adhesion assay with an IC₅₀ of less than about 10nM.

In Vivo Activities

Certain compounds of the invention exhibit in vivo biologicalactivities, such as those described herein in Examples 39-41.

Synthesis of the Compounds of the Invention

The compounds of the invention can be prepared using methods known inthe art of organic synthesis and those described herein (see, e.g.,Examples 1 to 13). The starting materials and various intermediates maybe obtained from commercial sources, prepared from commerciallyavailable compounds, and/or prepared using known synthetic methods. Forexample, the compounds of the invention, as well as all intermediates,can be synthesized by known processes using either solution or solidphase techniques. Exemplary procedures for preparing compounds of theinvention are outlined in the following schemes.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. Suitableprotecting groups for various functional groups as well as suitableconditions for protecting and deprotecting particular functional groupsare well known in the art. For example, numerous protecting groups aredescribed in T. W. Greene and P. G. M. Wuts, Protecting Groups inOrganic Synthesis, Third Edition, Wiley, New York, 1999, and referencescited therein.

In one example, the compounds of the invention are prepared using aprocedure outlined in Scheme 1, below:

In Scheme 1, the amine (a) is coupled to the dimethyl2-(3-methoxyallylidene)malonate (b). The resulting malonate derivative(c) is cyclized in the presence of a base, such as NaH, to give the2-oxo-1,2-dihydropyridine analog (d). The carboxylic acid group of (d)is utilized to form an amide bond with an amine derivative incorporatingthe ring Cy, thereby forming a compound of the invention.

Compounds of formula (a) in Scheme 1 can be synthesized from thecorresponding ketone (f) by reaction with a hydroxylamine and subsequentreduction of the resulting oxime (g), e.g., as outlined in Scheme 2,below.

Alternatively compounds of formula (a) can be synthesized from thecorresponding ketone as outlined in Scheme 3a, below.

In Scheme 3a, X is Br or I. R¹ and R² are as defined herein above. InScheme 3, the halogen-substituted ketone (h) is first converted to thealkyl-oxime (i). The halogen can then be replaced with another group(e.g., an alkyl group, an amine, or an alkoxy group) using a variety ofknown chemistries. In one example, the halogen X (e.g., Br) is replacedwith an alkyl or cycloalkyl group using a Suzuki or Suzuki-typereaction, e.g., employing a boronic acid reagent (j) to afford analog(k). The oxime can subsequently be reduced to the corresponding amine(l).

In another example, the halogen of compound (h) is replaced with anamino group, e.g., using a Buchwald or Buchwald-type reaction asoutlined in Scheme 3b, below.

In Scheme 3b, X is Br or I and R^(a) and R^(b) are members independentlyselected from H, substituted or unsubstituted alkyl (e.g., C₁-C₁₀alkyl), substituted or unsubstituted heteroalkyl (e.g., 2- to10-membered heteroalkyl), substituted or unsubstituted aryl, substitutedor unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl(e.g., C₃-C₁₀ cycloalkyl) and substituted or unsubstitutedheterocycloalkyl (e.g., 3- to 10-membered heterocycloalkyl). R^(a) andR^(b), together with the nitrogen atom, to which they are attached, areoptionally joined to form a 4 to 7-membered heterocyclic ring. Exemplaryheterocyclic rings include piperazinyl (e.g., N-methyl-piperazinyl) andmorpholinyl groups. In Scheme 3b, the halogen X (e.g., Br) of the oxime(i) is replaced with an amine (m) using a metal catalyst, such as atransition metal catalyst to afford the amine (n). The oxime moiety of(n) can then be reduced to the amine forming analog (O). Catalystsuseful for the above transformation are known to those of skill in theart. It is well within the capabilities of a skilled person to select asuitable catalyst. Typically, the Buchwald or Buchwald-type reactionwill be palladium-catalyzed. However, other transition metal catalystscan also be used. For example, the starting material can be heatedtogether with a palladium catalyst, a palladium ligand and a base. Inone example, the catalyst is palladium acetate (e.g., Pd(OAc)₂) or apalladium phosphine, such as triphenyl phosphine, Pd(PPh₃)₄. Thereaction (i.e., replacing a halogen with an amino group) can also beperformed later in the synthesis, e.g., after the pyridone ring hasalready been formed.

Alternatively, in Scheme 3a or 3b, X is F, which can be replaced with athiol or an alkylthio (thioether) group as outlined in Scheme 4, below.

In Scheme 4, the fluoro-substituted ketone (p) is first converted to thealkyl-oxime (q). The halogen can then be replaced with a thioether groupby reacting compound (q) with an alkyl thiolate (r), wherein R^(a) isalkyl (e.g., C₁-C₄ alkyl, such as methyl or ethyl), to afford thethioether (s), which can be further reduced to the amine (t). Thethioether group can optionally be oxidized to a sulfonyl group, e.g.,later in the synthesis, using an oxidizing agent, such as mCPBA, asoutlined in Scheme 5, below.

Useful ketone analogs, such as (f), (h) and (p) in Schemes 2, 3 and 4,respectively, can be prepared using a procedure outlined in Scheme 6,below.

In Scheme 6, condensation of the aldehyde (u) with malonic acid or amalonic acid derivative (v) affords the unsaturated analog (w). Thedouble bond can be reduced to afford the saturated derivative (x).Cyclization of (x) to the dihydroindenone (y) can be accomplished, e.g.,using a dehydrating agent, such as P₂O₅.

In another example, amino-substituted compounds of the invention areprepared using a procedure outlined in Scheme 7, below.

In Scheme 7, Cy and R¹ are defined as herein above. In Scheme 7, thenitro-substituted ketone (z) is first reduced to the correspondingalcohol (aa), which is coupled to the pyridone (bb) thereby forminganalog (cc). The nitro group of (cc) is then reduced to thecorresponding amine (dd).

The amino analog (dd) of Scheme 7 can be further converted to thesecondary amine (ee) or the tertiary amine (ff). Alternatively the aminogroup of (dd) can be replaced with a halogen to afford thehalogen-substituted analog (gg) or a nitrile (CN) group, e.g., viaSandmeyer or Sandmeyer-type reactions. These conversions are summarizedin Scheme 8, below.

In Scheme 8, X is halogen (e.g., Cl, Br or I) and R^(c) and R^(d) aremembers independently selected from H, substituted or unsubstitutedalkyl (e.g., C₁-C₁₀ alkyl), substituted or unsubstituted heteroalkyl(e.g., 2- to 10-membered heteroalkyl), substituted or unsubstitutedaryl, substituted or unsubstituted heteroaryl, substituted orunsubstituted cycloalkyl (e.g., C₃-C₁₀ cycloalkyl) and substituted orunsubstituted heterocycloalkyl (e.g., 3- to 10-memberedheterocycloalkyl). R^(c) and R^(d), together with the nitrogen atom, towhich they are attached, are optionally joined to form a 4 to 7-memberedheterocyclic ring.

In another example, the compounds of the invention are prepared using aprocedure outlined in Scheme 9, below. Phenolic hydroxyl groups atvarious positions on the molecule can be used to attach a linker moiety,such as a protected (e.g., Boc-protected) amino-ethylene or -propyleneglycol moiety, e.g., via a Mitsunobu or Mitsunobu-type reaction. Aftercoupling, the amino protecting group can be removed to afford a primaryamino group, which can be used to covalently attach the compound toanother molecule, such as a second compound of the invention (to createa dimeric inhibitor) or a branched linker molecule (e.g., glycerol) tocreate a conjugate. The linker can be used to covalently attach one ormore additional compounds of the invention to create a multimericconjugate. Dimeric and multimeric integrin antagonists and theirsyntheses have been described e.g., in WO2006/010054 and WO2005/070921,the disclosures of which are incorporated herein by reference in theirentirety.

Further, compounds of Formula (Ia), Formula (Ia′), Formula (Ib) andFormula (Ic) can be synthesized following the procedures outlined in theabove schemes using the starting compounds shown in Scheme 10. Suchcompounds shown in Scheme 10 can be purchased from commercial sources orsynthesized using the methods disclosed herein or methods known to oneof skill in the art.

Pharmaceutical Compositions

The invention further provides pharmaceutical compositions including acompound of the invention, e.g., those of Formulae (I) to (XV), and apharmaceutically acceptable carrier. Pharmaceutically acceptablecarriers are described herein. A pharmaceutical composition of theinvention may include one or more compounds of the invention inassociation with one or more pharmaceutically acceptable carriers andoptionally other active ingredients.

The compounds of the invention may be administered orally, topically,parenterally, by inhalation or spray or rectally in dosage unitformulations containing at least one pharmaceutically acceptablecarrier. The term “carrier” includes adjuvants, diluents, excipients andvehicles. The term “parenteral” as used herein includes percutaneous,subcutaneous, intravascular (e.g., intravenous), intramuscular, orintrathecal injection or infusion techniques and the like. Thepharmaceutical compositions containing compounds of the invention may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsion, hard or soft capsules, or syrups or elixirs.

Compositions intended for oral use may be prepared according to anymethod known to the art for the manufacture of pharmaceuticalcompositions and such compositions may contain one or more agentsselected from the group consisting of sweetening agents, flavoringagents, coloring agents and preservative agents in order to providepharmaceutically elegant and palatable preparations. Tablets contain theactive ingredient in admixture with non-toxic pharmaceuticallyacceptable excipients that are suitable for the manufacture of tablets.These excipients may be for example, inert diluents, such as calciumcarbonate, sodium carbonate, lactose, calcium phosphate or sodiumphosphate; granulating and disintegrating agents, for example, cornstarch, or alginic acid; binding agents, for example starch, gelatin oracacia, and lubricating agents, for example magnesium stearate, stearicacid or talc. The tablets may be uncoated or they may be coated by knowntechniques. In some cases such coatings may be prepared by knowntechniques to delay disintegration and absorption in thegastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonosterate or glyceryl distearate may be employed.

Formulations for oral use may also be presented as hard gelatincapsules, wherein the active ingredient is mixed with an inert soliddiluent, for example, calcium carbonate, calcium phosphate or kaolin, oras soft gelatin capsules wherein the active ingredient is mixed withwater or an oil medium, for example peanut oil, liquid paraffin or oliveoil. Formulations for oral use may also be presented as lozenges.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydropropyl-methylcellulose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example, lecithin, or condensation products of an alkylene oxidewith fatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientsin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents and flavoring agents may beadded to provide palatable oral preparations. These compositions may bepreserved by the addition of an anti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents orsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

Pharmaceutical compositions of the invention may also be in the form ofoil-in-water emulsions. The oily phase may be a vegetable oil or amineral oil or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitol,anhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol, glucose or sucrose. Suchformulations may also contain a demulcent, a preservative and flavoringand coloring agents. The pharmaceutical compositions may be in the formof a sterile injectable aqueous or oleaginous suspension. Thissuspension may be formulated according to the known art using thosesuitable dispersing or wetting agents and suspending agents that havebeen mentioned above. The sterile injectable preparation may also be asterile injectable solution or suspension in a non-toxic parentallyacceptable diluent or solvent, for example as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose any bland fixed oilmay be employed including synthetic mono- or diglycerides. In addition,fatty acids such as oleic acid find use in the preparation ofinjectables.

The compounds of the invention may also be administered in the form ofsuppositories, e.g., for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient that is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials include cocoa butter andpolyethylene glycols.

Compounds of the invention may be administered parenterally in a sterilemedium. The compound, depending on the vehicle and concentration used,can either be suspended or dissolved in the vehicle. Advantageously,adjuvants such as local anesthetics, preservatives and buffering agentscan be dissolved in the vehicle.

For disorders of the eye or other external tissues, e.g., mouth andskin, the formulations are preferably applied as a topical gel, spray,ointment or cream, or as a scleral suppository, containing the activeingredients in a total amount of, for example, 0.075 to 30% w/w,preferably 0.2 to 20% w/w and most preferably 0.4 to 15% w/w. Whenformulated in an ointment, the active ingredients may be employed witheither paraffinic or a water-miscible ointment base.

Alternatively, the active ingredients may be formulated in a cream withan oil-in-water cream base. If desired, the aqueous phase of the creambase may include, for example at least 30% w/w of a polyhydric alcoholsuch as propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerol,polyethylene glycol and mixtures thereof. The topical formulation maydesirably include a compound, which enhances absorption or penetrationof the active ingredient through the skin or other affected areas.Examples of such dermal penetration enhancers include dimethylsulfoxideand related analogs. The compounds of this invention can also beadministered by a transdermal device. Preferably topical administrationwill be accomplished using a patch either of the reservoir and porousmembrane type or of a solid matrix variety. In either case, the activeagent is delivered continuously from the reservoir or microcapsulesthrough a membrane into the active agent permeable adhesive, which is incontact with the skin or mucosa of the recipient. If the active agent isabsorbed through the skin, a controlled and predetermined flow of theactive agent is administered to the recipient. In the case ofmicrocapsules, the encapsulating agent may also function as themembrane. The transdermal patch may include the compound in a suitablesolvent system with an adhesive system, such as an acrylic emulsion, anda polyester patch. The oily phase of the emulsions of this invention maybe constituted from known ingredients in a known manner. While the phasemay comprise merely an emulsifier, it may comprise a mixture of at leastone emulsifier with a fat or oil or with both a fat and an oil.Preferably, a hydrophilic emulsifier is included together with alipophilic emulsifier, which acts as a stabilizer. It is also preferredto include both an oil and a fat. Together, the emulsifier(s) with orwithout stabilizer(s) make-up the so-called emulsifying wax, and the waxtogether with the oil and fat make up the so-called emulsifying ointmentbase, which forms the oily, dispersed phase of the cream formulations.Emulsifiers and emulsion stabilizers suitable for use in the formulationof the invention include Tween 60, Span 80, cetostearyl alcohol,myristyl alcohol, glyceryl monostearate, and sodium lauryl sulfate,among others. The choice of suitable oils or fats for the formulation isbased on achieving the desired cosmetic properties, since the solubilityof the active compound in most oils likely to be used in pharmaceuticalemulsion formulations is very low. Thus, the cream should preferably bea non-greasy, non-staining and washable product with suitableconsistency to avoid leakage from tubes or other containers. Straight orbranched chain, mono- or dibasic alkyl esters such as di-isoadipate,isocetyl stearate, propylene glycol diester of coconut fatty acids,isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate,2-ethylhexyl palmitate or a blend of branched chain esters may be used.These may be used alone or in combination depending on the propertiesrequired. Alternatively, high melting point lipids such as white softparaffin and/or liquid paraffin or other mineral oils can be used.

Formulations suitable for topical administration to the eye also includeeye drops wherein the active ingredients are dissolved or suspended insuitable carrier, especially an aqueous solvent for the activeingredients. The anti-inflammatory active ingredients are preferablypresent in such formulations in a concentration of 0.5 to 20%,advantageously 0.5 to 10% and particularly about 1.5% w/w. Fortherapeutic purposes, the active compounds of this combination inventionare ordinarily combined with one or more adjuvants appropriate to theindicated route of administration. The compounds may be admixed withlactose, sucrose, starch powder, cellulose esters of alkanoic acids,cellulose alkyl esters, talc, stearic acid, magnesium stearate,magnesium oxide, sodium and calcium salts of phosphoric and sulfuricacids, gelatin, acacia gum, sodium alginate, polyvinylpyrrolidone,and/or polyvinyl alcohol, and then tableted or encapsulated forconvenient administration. Such capsules or tablets may contain acontrolled-release formulation as may be provided in a dispersion ofactive compound in hydroxypropylmethyl cellulose. Formulations forparenteral administration may be in the form of aqueous or non-aqueousisotonic sterile injection solutions or suspensions. These solutions andsuspensions may be prepared from sterile powders or granules having oneor more of the carriers or diluents mentioned for use in theformulations for oral administration. The compounds may be dissolved inwater, polyethylene glycol, propylene glycol, ethanol, corn oil,cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride,and/or various buffers. Other adjuvants and modes of administration arewell and widely known in the pharmaceutical art.

Dosage levels of the order of from about 0.005 mg to about 80 mg perkilogram of body weight per day are useful in the treatment of thediseases and conditions described herein (e.g., about 0.35 mg to about5.6 g per human patient per day, based on an average adult person weightof 70 kg). The amount of active ingredient that may be combined with thecarrier materials to produce a single dosage form will vary dependingupon the host treated and the particular mode of administration. Dosageunit forms will generally contain between from about 1 mg to about 500mg of an active ingredient. The daily dose can be administered in one tofour doses per day. In the case of skin conditions, it may be preferableto apply a topical preparation of compounds of this invention to theaffected area one to four times a day.

Formulations suitable for inhalation or insufflation include solutionsand suspensions in pharmaceutically acceptable aqueous or organicsolvents, or mixtures thereof, and powders. The liquid or solidcompositions may contain suitable pharmaceutically acceptable excipientsas describe above. The compositions may be administered by oral or nasalrespiratory route for local or systemic effect. Compositions may benebulized by use of inert gases or vaporized, and breathed directly fromthe nebulizing/vaporizing device or the nebulizing device may beattached to a facemask tent or intermittent positive pressure-breathingmachine.

It will be understood, however, that the specific dose level for anyparticular patient will depend upon a variety of factors including theactivity of the specific compound employed, the age, body weight,general health, sex, diet, time of administration, route ofadministration, and rate of excretion, drug combination and the severityof the particular disease undergoing therapy.

For administration to non-human animals, the composition may also beadded to the animal feed or drinking water. It may be convenient toformulate the animal feed and drinking water compositions so that theanimal takes in a therapeutically appropriate quantity of thecomposition along with its diet. It may also be convenient to presentthe composition as a premix for addition to the feed or drinking water.

Methods

The conjugates of this invention are anticipated to exhibit inhibition,in vivo, of adhesion of leukocytes to endothelial cells mediated byintegrins, alpha4beta7 and VLA-4 by competitive binding to the integrin.Preferably, the compounds of this invention can be used in intravenousformulations for the treatment of diseases mediated by such integrins ordiseases involving leukocyte adhesion. Such diseases includeinflammatory diseases in mammalian patients such as asthma, Alzheimer'sdisease, atherosclerosis, AIDS dementia, diabetes (including acutejuvenile onset diabetes), inflammatory bowel disease (includingulcerative colitis and Crohn's disease), multiple sclerosis (MS),idiopathic pulmonary fibrosis (IPF; also referred to as cryptogenicfibrosing alveolitis), rheumatoid arthritis (RA), tissuetransplantation, tumor metastasis, liquid tumors, meningitis,encephalitis, stroke, and other cerebral traumas, nephritis, retinitis,atopic dermatitis, psoriasis, myocardial ischemia and acuteleukocyte-mediated lung injury such as that which occurs in adultrespiratory distress syndrome. The compounds and pharmaceuticalformulations of the present invention are especially useful in thetreatment of inflammatory diseases of the stomach/gut/intestines, suchas inflammatory bowel diseases (IBD), including Crohn's disease andulcerative colitis.

Inflammatory bowel disease refers to the group of disorders that causethe intestines to become inflamed, generally manifested with symptomsincluding abdominal cramps and pain, diarrhea, weight loss andintestinal bleeding. IBD is a collective term for two similar diseasestermed ulcerative colitis (UC) and Crohn's disease (CD).

Crohn's disease is a chronic autoimmune disorder that results ininflammation of the gastrointestinal (GI) tract. Although any area ofthe GI tract may be involved, CD most commonly affects the smallintestine and/or colon. In Crohn's disease, all layers of the intestinemay be involved, and there can be normal healthy bowel in betweenpatches of diseased bowel. CD is associated with fibrosis, stenosis andfissuring, fistulae between disease tracts and adjacent structures(i.e., bladder, other bowel segments, skin) and abscess. CD patients aretypically present with diarrhea, abdominal pain and weight loss. Theabdominal pain usually is insidious and may be associated with a tender,inflammatory mass. Fever, weight loss, stomatitis, perianal fistulaeand/or fissure, arthritis, and erythema nodosum are all commonly seen.There is considerable morbidity associated with CD, particularly inpatients with disease not controlled by currently available drugs. Up to75% of patients with moderate to severe disease require surgery and upto 75% to these patients will experience post surgical diseaserecurrence within 10 years and up to 50% will undergo a repeat surgerywithin 20 years. This high rate of recurrence indicates a need for both,new effective treatments for the active disease and maintenance ofdisease remission.

Ulcerative colitis is a chronic, episodic, inflammatory disease of thelarge intestine and rectum characterized by bloody diarrhea. Ulcerativecolitis is an inflammatory response limited largely to the colonicmucosa and submucosa. Ulcerative colitis can be categorized according tolocation: “proctitis” involves only the rectum, “proctosigmoiditis”affects the rectum and sigmoid colon, “left-sided colitis” encompassesthe entire left side of the large intestine, “pancolitis” inflames theentire colon. Lymphocytes and macrophages are numerous in lesions ofinflammatory bowel disease and may contribute to inflammatory injury. Anexemplary animal model of inflammatory bowel disease (IBD) is carriedout with HLA-B27 transgenic rats. These rats overexpress the humanHLA-B27 molecule (heavy chain and beta globulin gene) that is associatedwith spondyloarthropathies, a group of inflammatory conditions affectingthe skeleton. Prior to onset of skeletal inflammatory changes theseanimals develop non-granulomatous inflammation in the small intestineand diffuse crypt abscesses on the colon, a pathology that is similar tothat of Crohn's Disease in humans. Efficacy studies can be performed inthe HLA-B27 transgenic rat IBD model with compounds of this invention,e.g., as described herein.

Asthma is a disease characterized by increased responsiveness of thetracheobronchial tree to various stimuli potentiating paroxysmalconstriction of the bronchial airways. The stimuli cause release ofvarious mediators of inflammation from IgE-coated mast cells includinghistamine, eosinophilic and neutrophilic chemotactic factors,leukotrines, prostaglandin and platelet activating factor. Release ofthese factors recruits basophils, eosinophils and neutrophils, whichcause inflammatory injury. Animal models for the in vivo study of asthmainclude the rat asthma model, the mouse asthma model and the sheepmodel, e.g., those described herein.

Atherosclerosis is a disease of arteries (e.g., coronary, carotid, aortaand iliac). The basic lesion, the atheroma, consists of a raised focalplaque within the intima, having a core of lipid and a covering fibrouscap. Atheromas compromise arterial blood flow and weaken affectedarteries. Myocardial and cerebral infarcts are a major consequence ofthis disease. Macrophages and leukocytes are recruited to atheromas andcontribute to inflammatory injury.

Rheumatoid arthritis is a chronic, relapsing inflammatory disease thatprimarily causes impairment and destruction of joints. Rheumatoidarthritis usually first affects the small joints of the hands and feetbut then may involve the wrists, elbows, ankles and knees. The arthritisresults from interaction of synovial cells with leukocytes thatinfiltrate from the circulation into the synovial lining of the joints.See e.g., Paul, Immunology (3d ed., Raven Press, 1993). Over time, boneerosion, destruction of cartilage, and complete loss of joint integritycan occur. Eventually, multiple organ systems may be affected.

Joint damage in rheumatoid arthritis begins with the proliferation ofsynovial macrophages and fibroblasts after a triggering incident,possibly autoimmune or infectious. Lymphocytes infiltrate perivascularregions, and endothelial cells proliferate. Neovascularization thenoccurs. Blood vessels in the affected joint become occluded with smallclots of inflammatory cells. Over time, inflamed synovial tissue beginsto grow irregularly, forming invasive pannus tissue. Pannus invades anddestroys cartilage and bone. Multiple cytokines, interleukins,proteinases, and growth factors are released, causing further jointdestruction and the development of systemic complications. See,Firestein G. S. Etiology and pathogenesis of rheumatoid arthritis, RuddyS, Harris E D, Sledge C B, Kelley W N, eds. Kelley's Textbook ofRheumatology, 7th ed. Philadelphia: W.B. Saunders, 2005:996-1042. Animalmodels for the study of rheumatoid arthritis include Adjuvant InducedArthritis (“AIA”) and Collagen Induced Arthritis (“CIA”).

Another indication for the compounds of this invention is in thetreatment of organ or graft rejection mediated by VLA-4. Over recentyears there has been a considerable improvement in the efficiency ofsurgical techniques for transplanting tissues and organs such as skin,kidney, liver, heart, lung, pancreas and bone marrow. Perhaps theprincipal outstanding problem is the lack of satisfactory agents forinducing immunotolerance in the recipient to the transplanted allograftor organ. When allogeneic cells or organs are transplanted into a host(i.e., the donor and donee are different individuals from the samespecies), the host immune system is likely to mount an immune responseto foreign antigens in the transplant (host-versus-graft disease)leading to destruction of the transplanted tissue. CD8⁺ cells, CD4 cellsand monocytes are all involved in the rejection of transplant tissues.Compounds of this invention which bind to alpha-4 integrin are useful,inter alia, to block alloantigen-induced immune responses in the doneethereby preventing such cells from participating in the destruction ofthe transplanted tissue or organ. See, e.g., Paul et al., TransplantInternational 9, 420-425 (1996); Georczynski et al., Immunology 87,573-580 (1996); Georcyznski et al., Transplant. Immunol. 3, 55-61(1995); Yang et al., Transplantation 60, 71-76 (1995); Anderson et al.,APMIS 102, 23-27 (1994).

A related use for compounds of this invention, which bind to VLA-4 is inmodulating the immune response involved in “graft versus host” disease(“GVHD”). See e.g., Schlegel et al., J. Immunol. 155, 3856-3865 (1995).GVHD is a potentially fatal disease that occurs when immunologicallycompetent cells are transferred to an allogeneic recipient. In thissituation, the donor's immunocompetent cells may attack tissues in therecipient. Tissues of the skin, gut epithelia and liver are frequenttargets and may be destroyed during the course of GVHD. The diseasepresents an especially severe problem when immune tissue is beingtransplanted, such as in bone marrow transplantation; but less severeGVHD has also been reported in other cases as well, including heart andliver transplants. The therapeutic agents of the present invention areused, inter alia, to block activation of the donor T-cells therebyinterfering with their ability to lyse target cells in the host.

Appropriate in vivo models for demonstrating efficacy in treatinginflammatory conditions include EAE (experimental autoimmuneencephalomyelitis) in mice, rats, guinea pigs or primates, as well asother inflammatory models dependent upon α₄ integrins.

A further use of the compounds of this invention is inhibiting tumormetastasis. Several tumor cells have been reported to express VLA-4 andcompounds, which bind VLA-4 block adhesion of such cells to endothelialcells. Steinback et al., Urol. Res. 23, 175-83 (1995); Orosz et al.,Int. J. Cancer 60, 867-71 (1995); Freedman et al., Leuk. Lymphoma 13,47-52 (1994); Okahara et al., Cancer Res. 54, 3233-6 (1994).

A further use of the compounds of this invention is in treating multiplesclerosis. Multiple sclerosis is a progressive neurological autoimmunedisease that is thought to be the result of an autoimmune reaction inwhich certain leukocytes attack and initiate the destruction of myelin,the insulating sheath covering nerve fibers. In an animal model formultiple sclerosis, murine monoclonal antibodies directed against VLA-4have been shown to block the adhesion of leukocytes to the endothelium,and thus prevent inflammation of the central nervous system andsubsequent paralysis in the animals.¹⁶

The most common demyelinating disease is multiple sclerosis, but manyother metabolic and inflammatory disorders result in deficient orabnormal myelination. MS is a chronic neurologic disease, which appearsin early adulthood and progresses to a significant disability in mostcases. There are approximately 350,000 cases of MS in the United Statesalone. Outside of trauma, MS is the most frequent cause of neurologicdisability in early to middle adulthood.

MS is characterized by chronic inflammation, demyelination and gliosis(scarring). Demyelination may result in either negative or positiveeffects on axonal conduction. Positive conduction abnormalities includeslowed axonal conduction, variable conduction block that occurs in thepresence of high- but not low-frequency trains of impulses or completeconduction block. Positive conduction abnormalities include ectopicimpulse generation, spontaneously or following mechanical stress andabnormal “cross-talk” between demyelinated exons.

T-cells reactive against myelin proteins, either myelin basic protein(MBP) or myelin proteolipid protein (PLP) have been observed to mediateCNS inflammation in experimental allergic encephalomyelitis. Patientshave also been observed as having elevated levels of CNS immunoglobulin(Ig). It is further possible that some of the tissue damage observed inMS is mediated by cytokine products of activated T cells, macrophages orastrocytes.

Today, 80% patients diagnosed with MS live 20 years after onset ofillness. Therapies for managing MS include: (1) treatment aimed atmodification of the disease course, including treatment of acuteexacerbation and directed to long-term suppression of the disease; (2)treatment of the symptoms of MS; (3) prevention and treatment of medicalcomplications; and (4) management of secondary personal and socialproblems.

The onset of MS may be dramatic or so mild as to not cause a patient toseek medical attention. The most common symptoms include weakness in oneor more limbs, visual blurring due to optic neuritis, sensorydisturbances, diplopia and ataxia. The course of disease may bestratified into three general categories: (1) relapsing MS, (2) chronicprogressive MS, and (3) inactive MS. Relapsing MS is characterized byrecurrent attacks of neurologic dysfunction. MS attacks generally evolveover days to weeks and may be followed by complete, partial or norecovery. Recovery from attacks generally occurs within weeks to severalmonths from the peak of symptoms, although rarely some recovery maycontinue for 2 or more years.

Chronic progressive MS results in gradually progressive worseningwithout periods of stabilization or remission. This form develops inpatients with a prior history of relapsing MS, although in 20% ofpatients, no relapses can be recalled. Acute relapses also may occurduring the progressive course.

A third form is inactive MS. Inactive MS is characterized by fixedneurologic deficits of variable magnitude. Most patients with inactiveMS have an earlier history of relapsing MS.

Disease course is also dependent on the age of the patient. For example,favourable prognostic factors include early onset (excluding childhood),a relapsing course and little residual disability 5 years after onset.By contrast, poor prognosis is associated with a late age of onset(i.e., age 40 or older) and a progressive course. These variables areinterdependent, since chronic progressive MS tends to begin at a laterage that relapsing MS. Disability from chronic progressive MS is usuallydue to progressive paraplegia or quadriplegia (paralysis) in patients.In one aspect of the invention, patients will preferably be treated whenthe patient is in remission rather than in a relapsing stage of thedisease.

Short-term use of either adrenocorticotropic hormone or oralcorticosteroids (e.g., oral prednisone or intravenousmethylprednisolone) is the only specific therapeutic measure fortreating patients with acute exacerbation of MS.

Newer therapies for MS include treating the patient with interferonbeta-1b, interferon beta-1a, and Copaxone® (formerly known as copolymer1). These three drugs have been shown to significantly reduce therelapse rate of the disease. These drugs are self-administeredintramuscularly or subcutaneously.

However, none of the current treatment modalities inhibit demyelination,let alone promotes or allows spontaneous remyelination or reducesparalysis. One aspect of the invention contemplates treating MS withagents disclosed herein either alone or in combination with otherstandard treatment modalities.

Radiation also can induce demyelination. Central nervous system (CNS)toxicity due to radiation is believed to be cause by (1) damage tovessel structures, (2) deletion of oligodendrocyte-2 astrocyteprogenitors and mature oligodendrocytes, (3) deletion of neural stemcell populations in the hippocampus, cerebellum and cortex, andgeneralized alterations of cytokine expression. Most radiation damageresults from radiotherapies administered during the treatment of certaincancers. See for review Belka et al., 2001 Br. J. Cancer 85: 1233-9.However, radiation exposure may also be an issue for astronauts(Hopewell, 1994 Adv. Space Res. 14: 433-42) as well as in the event ofexposure to radioactive substances.

Pharmaceutical compositions of the invention are suitable for use in avariety of drug delivery systems. Suitable formulations for use in thepresent invention are found in Remington's Pharmaceutical Sciences, MacePublishing Company, Philadelphia, Pa., 17th ed. (1985), the disclosureof which is incorporated herein in its entirety.

The amount administered to the patient will vary depending upon what isbeing administered, the purpose of the administration, such asprophylaxis or therapy, the state of the patient, the manner ofadministration, and the like. In therapeutic applications, compositionsare administered to a patient already suffering from a disease in anamount sufficient to cure or at least partially arrest the symptoms ofthe disease and its complications. An amount adequate to accomplish thisis defined as “therapeutically effective dose.” Amounts effective forthis use will depend on the disease condition being treated as well asby the judgment of the attending clinician depending upon factors suchas the severity of the inflammation, the age, weight and generalcondition of the patient, and the like, with reference to theappropriate animal model data, such as that provided herein. Methods forestimating appropriate human dosages, based on such data, are known inthe art. (see, for example, Wagner, J. G. Pharmacokinetics for thePharmaceutical Scientist. Technomic, Inc., Lancaster, Pa. 1993).

The compositions administered to a patient are in the form ofpharmaceutical compositions described above. These compositions may besterilized by conventional sterilization techniques, or may be sterilefiltered. The resulting aqueous solutions may be packaged for use as is,or lyophilized, the lyophilized preparation being combined with asterile aqueous carrier prior to administration.

Compounds having the desired biological activity may be modified asnecessary to provide desired properties such as improved pharmacologicalproperties (e.g., in vivo stability, bio-availability), or the abilityto be detected in diagnostic applications. Stability can be assayed in avariety of ways such as by measuring the half-life of the proteinsduring incubation with peptidases or human plasma or serum. A number ofsuch protein stability assays have been described (see, e.g., Verhoef etal., Eur. J. Drug Metab. Pharmacokinet, 1990, 15(2):83-93).

Inflammatory diseases that are included for treatment by thecompositions, compounds and methods disclosed herein include generallyconditions relating to demyelination. Histologically, myelinabnormalities are either demyelinating or dysmyelinating. Demyelinationimplies the destruction of myelin. Dysmyelination refers to defectiveformation or maintenance of myelin resulting from dysfunction of theoligodendrocytes. Preferably, the compositions and methods disclosedherein are contemplated to treat diseases and conditions relating todemyelination and aid with remyelination. Additional diseases orconditions contemplated for treatment include meningitis, encephalitis,and spinal cord injuries and conditions generally which inducedemyelination as a result of an inflammatory response.

The compositions, compounds and cocktails disclosed herein arecontemplated for use in treating conditions and diseases associated withdemyelination. Diseases and conditions involving demyelination include,but are not limited to, multiple sclerosis, congenital metabolicdisorders (e.g., phenylketonuria (PKU), Tay-Sachs disease, Niemann-Pickdisease, Gaucher's disease, Hurler's syndrome, Krabbe's disease andother leukodystrophies that impact the developing sheath), neuropathieswith abnormal myelination (e.g., Guillain Barré, chronic immunedemyelinating polyneuropathy (CIDP), multifocal CIDP, Multifocal MotorNeuropathy (MMN), anti-MAG (Myelin-Associated Glycoprotein) syndrome,GALOP (Gait disorder, Autoantibody, Late-age, Onset, Polyneuropathy)syndrome, anti-sulfatide antibody syndrome, anti-GM2 antibody syndrome,POEMS (Polyneuropathy, Organomegaly, Endocrinopathy, M-Protein and Skinchanges) syndrome also known as Crow-Fukase Syndrome and Takatsukidisease, perineuritis, IgM anti-GD1b antibody syndrome), drug relateddemyelination (e.g., caused by the administration of chloroquine, FK506,perhexyline, procainamide, and zimeldine), other hereditarydemyelinating conditions (e.g., carbohydrate-deficient glycoprotein,Cockayne's syndrome, congenital hypomyelinating, congenital musculardystrophy, Farber's disease, Marinesco-Sjögren syndrome, metachromaticleukodystrophy, Pelizaeus-Merzbacher disease, Refsum disease, prionrelated conditions, and Salla disease) and other demyelinatingconditions (e.g., meningitis, encephalitis (also known as acutedisseminated encephalomyelitis, ADEM), or spinal cord injury) ordiseases.

There are various disease models that can be used to study thesediseases in vivo. For example, animal models include but are not limitedto:

TABLE 4 Disease Model Species EAE Mouse, rat, guinea pigMyelin-oligodendrocyte glycoprotein Rat (MOG) induced EAE TNF-αtransgenic model of Mouse demyelination

These conditions and diseases are also contemplated for palliative orameliorating treatments.

Compounds of this invention are also capable of binding or antagonizingthe actions of α₄β₁, and α₄β₇ integrins. Accordingly, compounds of thisinvention are also useful for preventing or reversing the symptoms,disorders or diseases induced by the binding of these integrins to theirrespective ligands.

In another aspect of the invention, the compounds and compositionsdescribed herein can be used to inhibit immune cell migration from thebloodstream to the central nervous system in the instance of, forexample, multiple sclerosis, or to areas which result ininflammatory-induced destruction of the myelin. Preferably, thesereagents inhibit immune cell migration in a manner that inhibitsdemyelination and that further may promote remyelination. The reagentsmay also prevent demyelination and promote remyelination of the centralnervous system for congenital metabolic disorders in which infiltratingimmune cells affect the development myelin sheath, mainly in the CNS.The reagents preferably also reduce paralysis when administered to asubject with paralysis induced by a demyelinating disease or condition.

Use of Compounds of the Invention in an In Vitro Assay

The invention further provides a method of using a compound of theinvention in an in vitro assay measuring binding of an α4β1 or α4β7integrin to an integrin ligand, such as fibronectin (FN), VCAM-1,osteopontin and MadCAM. For example, the compound of the invention canbe used as a reference compound in such assay in order to compare andevaluate the binding capacity of other test compounds (i.e., candidatemolecules) to bind the integrin, or its capacity to disrupt binding ofthe ligand to the integrin (i.e., competitive binding assay). The term“in vitro assay” includes cell-based (e.g., ex vivo) assays. Anexemplary cell-based in vitro assay measuring the capacity of testcompounds to inhibit binding of MadCAM to cell-surface alpha4beta7integrins is described herein. An exemplary assay comprises the steps of(i) binding the ligand (e.g., recombinant MadCAM-antibody conjugate) toa surface (e.g., a well-plate); (ii) contacting the ligand with a cellthat expresses the integrin (e.g. alpha4beta7) on its cell-surface inthe presence of a compound of the invention; and (iii) measuring theamount of cells bound to the surface. For example, the integrins on thesurface of the cells bind to the immobilized ligand. Such binding isinhibited by a compound of the invention as the compound binds to theintegrin and thus reduces binding of the integrin to the ligand.

The invention further provides a method of using a compound of theinvention in an in vitro assay measuring binding of the compound to anα4β1 or α4β7 integrin in the presence of a test molecule. For example,solubilized integrin or membrane preparations containing the integrincan be incubated with a test molecule in the presence of a compound ofthe invention labeled with a radioactive, colorimetric, fluorescent orother label, which can be used for detection. The test molecule competeswith the labeled compound for binding to the integrin. By measuring theamount of labeled compound bound to the integrin, the capability of thetest molecule to the integrin can be determined. Hence, the inventionfurther provides a method of using the compounds of the invention in anin vitro assay for identifying a candidate molecule capable of bindingto α4β1 or α4β7 integrin. A candidate molecule exhibits a detectablebinding activity in the respective assay (e.g., an IC₅₀ of not more than10 μM and preferably not more than 5 μM or not more than 1 μM).

In one example according to any of the above embodiments, the in vitroassay is a competitive binding assay.

The disclosures in this document of all articles and references,including patents, are incorporated herein by reference in theirentirety.

The invention is illustrated further by the following examples, whichare not to be construed as limiting the invention in scope or spirit tothe specific procedures described in them. Analogous structures andalternative synthetic routes within the scope of the invention will beapparent to those skilled in the art.

EXAMPLES General

Reagents and solvents obtained from commercial suppliers were usedwithout further purification unless otherwise stated. Thin layerchromatography was performed on precoated 0.25 mm silica gel plates (E.Merck, silica gel 60, F₂₅₄). Visualization was achieved using UVillumination or staining with phosphomolybdic acid, ninhydrin or othercommon staining reagents. Flash chromatography was performed usingeither a Biotage Flash 40 system and prepacked silica gel columns orhand packed columns (E. Merck silica gel 60, 230-400 mesh). PreparatoryHPLC was performed on a Varian Prepstar high performance liquidchromatograph. ¹H and ¹³C NMR spectra were recorded at 300 MHz and 75MHz, respectively, on a Varian Gemini or Bruker Avance spectrometer.Chemical shifts are reported in parts per million (ppm) downfieldrelative to tetramethylsilane (TMS) or to proton resonances resultingfrom incomplete deuteration of the NMR solvent (8 scale). Mass spectrawere recorded on an Agilent series 1100 mass spectrometer connected toan Agilent series 1100 HPLC.

Further, abbreviations used throughout have the following meanings:

-   -   μ=Micro    -   Ac=Acetate    -   bd=Broad doublet    -   BINAP=2,2′-Bis(diphenylphosphino)-1,1′-binaphthyl    -   BOC=tert-Butyloxycarbonyl    -   BSA=Bovine serum albumin    -   d=Doublet    -   DCM=Dichloromethane    -   dd=Double doublet    -   DEAD=Diethyl azodicarboxylate    -   DIAD=Diisopropyl azodicarboxylate    -   DIEA=Diisopropylethylamine    -   DME=Dimethylether    -   DMF=Dimethylformamide    -   DMSO=Dimethylsulfoxide    -   dppf=1,1′-Bis(diphenylphosphino)ferrocene    -   dppp=1,1-Bis(diphenylphosphino)methane    -   EDC=1-Ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride    -   eq=Equivalents    -   Et=Ethyl    -   FBS=fetal bovine serum    -   g=Grams    -   HBTU=2-(1H-Benzotriazol-1-yl)-1,1,3,3-tetramethyluronium        hexafluorophosphate    -   HEPES=4-2-Hydroxyethyl-1-piperazineethanesulfonic acid    -   HOBt=1-Hydroxybenzotriazole    -   HPLC=High performance liquid chromatography    -   hr=Hours    -   LAH=Lithium aluminum hydride    -   m=Multiplet    -   M=Molar    -   m-CPBA=meta-Chloroperoxybenzoic acid    -   MeOH=Methanol    -   mg=Milligrams    -   MHz=Megahertz    -   min=Minute    -   ml=Milliliters    -   mM=Millimolar    -   mmol=Millimole    -   MS (ESI)=Electrospray ionization mass spectrometry    -   N=Normal    -   NMR=Nuclear magnetic resonance    -   Pd/C=Palladium on carbon    -   Pd₂(dba)₃=Tris(dibenzylideneacetone)dipalladium(0)    -   Ph=Phenyl    -   psi=Pounds per square inch    -   q=Quartet    -   Ra-Ni=Raney-Nickel    -   rpm=Rotation per minute    -   RT=Room temperature    -   s=Singlet    -   t=Triplet    -   TFA=Trifluoroacetic acid    -   THF=Tetrahydrofuran    -   uL or μl=Microliters    -   w/v=Weight/volume=    -   δ=Chemical shift

Compound purity was typically determined by HPLC/MS analysis using avariety of analytical methods. Exemplary methods are described below.

Method [1]=20% [B]: 80% [A] to 70% [B]: 30% [A] gradient in 1.75 min,then hold, at 2 mL/min, where [A]=0.1% trifluoroacetic acid in water;[B]=0.1% trifluoroacetic acid in acetonitrile on a Phenomenex Luna C18(2) 4.6 mm×30 cm column, 3 micron packing, 210 nm detection, at 35° C.Method [2]=50% [B]: 50% [A] to 95% [B]: 5% [A] gradient in 2.5 min, thenhold, at 2 mL/min, where [A]=0.1% trifluoroacetic acid in water;[B]=0.1% trifluoroacetic acid in acetonitrile on a Phenomenex Luna C18(2) 4.6 mm×30 cm column, 3 micron packing, 210 nm detection, at 35° C.Method [3]=5% [B]: 95% [A] to 20% [B]: 80% [A] gradient in 2.5 min, thenhold, at 2 mL/min, where [A]=0.1% trifluoroacetic acid in water;[B]=0.1% trifluoroacetic acid in acetonitrile on a Phenomenex Luna C18(2) 4.6 mm×30 cm column, 3 micron packing, 210 nm detection, at 35° C.Method [4]=20% [B]: 80% [A] to 70% [B]: 30% [A] gradient in 2.33 min,then hold, at 1.5 mL/min, where [A]=0.1% trifluoroacetic acid in water;[B]=0.1% trifluoroacetic acid in acetonitrile on a Phenomenex Luna C18(2) 4.6 mm×30 cm column, 3 micron packing, 210 nm detection, at 35° C.Method [5]=50% [B]: 50% [A] to 95% [B]: 5% [A] gradient in 3.33 min,then hold, at 1.5 mL/min, where [A]=0.1% trifluoroacetic acid in water;[B]=0.1% trifluoroacetic acid in acetonitrile on a Phenomenex Luna C18(2) 4.6 mm×30 cm column, 3 micron packing, 210 nm detection, at 35° C.Method [6]=5% [B]: 95% [A] to 20% [B]: 80% [A] gradient in 3.33 min,then hold, at 1.5 mL/min, where [A]=0.1% trifluoroacetic acid in water;[B]=0.1% trifluoroacetic acid in acetonitrile on a Phenomenex Luna C18(2) 4.6 mm×30 cm column, 3 micron packing, 210 nm detection, at 35° C.Method [7]=20% [B]: 80% [A] to 70% [B]: 30% [A] gradient in 10.0 min,then hold, at 1.5 mL/min, where [A]=0.1% trifluoroacetic acid in water;[B]=0.1% trifluoroacetic acid in acetonitrile on a Phenomenex Luna C18(2) 4.6 mm×3 cm column, 3 micron packing, 210 nm detection, at 35° C.Method [8]=10% [B]: 90% [A] to 40% [B]: 60% [A] gradient in 10.0 min,then hold, at 1.5 mL/min, where [A]=0.1% trifluoroacetic acid in water;[B]=0.1% trifluoroacetic acid in acetonitrile on a Phenomenex Luna C18(2) 4.6 mm×3 cm column, 3 micron packing, 210 nm detection, at 35° C.Method [9]=23% [B]: 77% [A] to 30% [B]: 70% [A] gradient in 15.0 min,then hold, at 1.0 mL/min, where [A]=0.1% trifluoroacetic acid in water;[B]=0.1% trifluoroacetic acid in acetonitrile on a Zorbex SB-phenyl C182.1 mm×5 cm column, 5 micron packing, 210 nm detection, at 30° C.

Example 1 1.1. Synthesis of(R)-1-(2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(1) Protocol A

A 500 ml round-bottom flask was charged with 5.0 g (1.0 eq.)(R)-2,3-dihydro-1H-inden-1-amine and 7.5 g (1.0 eq.) dimethyl2-(3-methoxyallylidene)malonate in 120 ml dry THF. The mixture washeated at 65° C. under N₂ for 3 hrs. Then the solvent was removed invacuo, and the crude mixture was purified by silica gel flash columnchromatography to afford (R)-dimethyl2-(3-(2,3-dihydro-1H-inden-1-ylamino)allylidene)malonate as a yellowsolid (11.0 g) in 97% yield.

Protocol B

To a solution of 11.0 g (1.0 eq) (R)-dimethyl2-(3-(2,3-dihydro-1H-inden-1-ylamino)allylidene)malonate in 80 ml dryMeOH was slowly added 3.65 g (2.5 eq) NaH (60% dispersion in mineraloil) under N₂ protection. The reaction was stirred at room temperaturefor 10 minutes and was then heated at 60° C. After 20 minutes, to thereaction mixture were added 30 ml water. The resulting mixture washeated at 60° C. for an additional 20 minutes. The solvent was removedin vacuo. The residue was diluted with 50 ml water, and extracted withhexane (2×50 ml) to remove mineral oil. Then the aqueous mixture wasacidified with 2N HCl to pH 1-2 and extracted with ethyl acetate (2×100ml). The combined organic phase was washed with 100 ml brine, dried oversodium sulfate and concentrated in vacuo to afford(R)-1-(2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxylicacid as a white solid (8.7 g, yield 93%) which was used without furtherpurification.

Protocol C

To a solution of 7.8 g (1.0 eq.)(R)-1-(2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxylicacid in 100 ml DMF was added DIEA (10 ml, 2.5 eq.), HBTU (13.8 g, 1.2eq.) and 4-amine-pyridin (3.45 g, 1.2 eq.). The resulting mixture wasstirred at rt. overnight. The reaction was diluted with 300 ml water.The precipitate was collected by filtration, and washed with 2×50 mlwater. The solid was purified on short flash column in silica gel toprovide white solid 8.1 g (80%)(R)-1-(2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide.Retention time (min)=2.592, method [7], MS (ESI) 332.1 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.99 (s, 1H), 8.72 (d, 2H), 8.49 (d, 1H), 8.19 (d,2H), 7.68 (d, 1H), 7.36 (m, 2H), 7.33 (m, 1H), 7.24 (m, 1H), 7.15 (d,1H), 6.61 (t, 1H), 6.49-6.52 (m, 1H), 3.03-3.16 (m, 1H), 2.71-2.98 (m,1H), 2.66-2.69 (m, 1H), 2.06-2.14 (m, 1H).

1.2. Synthesis of Additional Compounds Using Protocols A, B and C

The following compounds were synthesized from appropriate startingmaterials using Protocols A, B and C, above or slightly modifiedversions thereof

1-(2,3-Dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(2)

Retention time (min)=2.757, method [7], MS (ESI) 332.1 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.98 (s, 1H), 8.71 (d, 2H), 8.48 (d, 1H), 8.18 (d,2H), 7.68 (d, 1H), 7.36 (m, 2H), 7.33 (m, 1H), 7.24 (t, 1H), 7.15 (d,1H), 6.61 (t, 1H), 6.49-6.52 (m, 1H), 3.01-3.19 (m, 1H), 2.71-2.98 (m,1H), 2.65-2.69 (m, 1H), 2.11-2.14 (m, 1H).

N-(4-Chlorophenyl)-1-(2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(3)

Retention time (min)=8.869, method [7], MS (ESI) 365.0 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.17 (s, 1H), 8.59 (dd, 1H), 7.74 (d, 2H), 7.28-7.39(m, 5H), 7.21 (dd, 1H), 7.19 (d, 1H), 6.61 (q, 1H), 6.39 (t, 1H),2.99-3.16 (m, 1H), 2.79-2.91 (m, 1H), 1.97-2.09 (m, 1H).

(S)-1-(2,3-Dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(4)

Retention time (min)=2.691, method [7], MS (ESI) 332.1 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.38 (s, 1H), 843-8.47 (m, 3H), 7.69 (d, 2H), 7.59(d, 1H), 7.40 (m, 2H), 7.35 (m, 1H), 7.24 (t, 1H), 7.15 (d, 1H),6.50-6.59 (m, 2H), 3.02-3.18 (m, 12H), 2.70-2.99 (m, 1H), 2.62-2.68 (m,1H), 2.02-2.14 (m, 1H).

(S)—N-(4-Chlorophenyl)-1-(2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(5)

Retention time (min)=8.464, method [7], MS (ESI) 365.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.18 (s, 1H), 8.58 (d, 1H), 7.74 (d, 2H), 7.28-7.39(m, 5H), 7.21 (d, 1H), 7.19 (d, 1H), 6.61 (q, 1H), 6.39 (t, 1H),2.91-3.18 (m, 1H), 2.79-2.89 (m, 1H), 2.05-2.09 (m, 1H).

(R)—N-(4-Chlorophenyl)-1-(2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(6)

Retention time (min)=9.461, method [7], MS (ESI) 365.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.16 (s, 1H), 8.60 (d, 1H), 7.74 (d, 2H), 7.28-7.39(m, 5H), 7.21 (d, 1H), 7.19 (d, 1H), 6.61 (q, 1H), 6.39 (t, 1H),2.91-3.16 (m, 1H), 2.79-2.89 (m, 1H), 2.05-2.09 (m, 1H).

N-(4-Chlorophenyl)-1-((1S,2R)-2-hydroxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(7)

Retention time (min)=7.443, method [7], MS (ESI) 381.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.06 (s, 1H), 8.51 (dd, 1H), 7.68 (d, 2H), 7.28-7.40(m, 5H), 7.22 (m, 1H), 7.13 (d, 1H), 6.44 (d, 1H), 6.37 (t, 1H), 4.97(m, 1H), 3.36 (dd, 1H), 3.05 (dd, 1H), 2.81 (d, 1H).

1-((1S,2R)-2-Hydroxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(8)

Retention time (min)=1.869, method [7], MS (ESI) 348.1 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.42 (s, 1H), 8.45-8.50 (m, 3H), 7.69 (d, 2H),7.49 (d, 1H), 7.23-7.39 (m, 3H), 7.08 (d, 1H), 6.57 (t, 1H), 6.48 (d,1H), 5.31 (d, 1H), 4.66 (m, 1H), 3.24 (m, 1H), 2.90-2.96 (d, 1H).

N-(4-Chlorophenyl)-1-((1R,2S)-2-hydroxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(9)

Retention time (min)=7.441, method [7], MS (ESI) 381.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.09 (s, 1H), 8.51 (d, 1H), 7.66 (d, 2H), 7.23-7.39(m, 5H), 7.22 (m, 1H), 7.13 (d, 1H), 6.44 (d, 1H), 6.37 (t, 1H), 4.99(m, 1H), 3.37 (dd, 1H), 3.05 (dd, 1H), 2.81 (d, 1H).

1-((1R,2S)-2-Hydroxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(10)

Retention time (min)=1.882, method [7], MS (ESI) 348.1 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.44 (s, 1H), 8.45-8.50 (m, 3H), 7.68 (d, 2H),7.49 (d, 1H), 7.23-7.39 (m, 3H), 7.08 (d, 1H), 6.57 (t, 1H), 6.48 (d,1H), 5.31 (d, 1H), 4.64 (m, 1H), 3.24 (m, 1H), 2.96 (d, 1H).

Example 2 2.1. Synthesis ofN-(4-chlorophenyl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(11) Protocol D

To a solution of 4-methoxy-2,3-dihydro-1H-inden-1-one (1.0 g, 1.0 eq.)in 25 ml MeOH was added NH₂OH.HCl (0.493 g, 1.15 eq.) and sodium acetate(0.597 g, 1.18 eq.). The resulting mixture was stirred at roomtemperature overnight. The reaction mixture was diluted with 100 mlice-water. The white precipitate was collected by filtration, washedwith 3×20 ml water and dried in vacuo to afford 1.08 g (yield 99%)4-methoxy-2,3-dihydro-1H-inden-1-one oxime which was used to next stepwithout further purification.

Protocol E

A solution of 4-methoxy-2,3-dihydro-1H-inden-1-one oxime (1.0 g, 1.0eq.) in 40 ml dry ether was cooled to −78° C. under N₂ atmosphere. ThenLiAlH₄ in ether solution (30 ml, 1M, 5.5 eq.) was added to reactiondropwise. The resulting mixture was stirred at −78° C. for 10 minutes,and gradually warmed to room temperature. Then the mixture was heated atrefluxing overnight. The reaction mixture was diluted with 50 ml etherand quenched by sequentially adding 1.2 ml water, 1.2 ml 15% NaOHaqueous solution and 3.4 ml water. The mixture was filtered throughcelite pad. The inorganic solid was rinsed with 3×20 ml ether. Theorganic phase was dried over Na₂SO₄, and evaporated in vacuo to afford aclear oil 0.5 g (yield 54.3%) 4-methoxy-2,3-dihydro-1H-inden-1-aminewhich was used to next step without further purification.

Following Protocols A, B and C, 4-methoxy-2,3-dihydro-1H-inden-1-aminewas converted toN-(4-chlorophenyl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide.Retention time (min)=9.577, method [7],

MS (ESI) 395.1 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.18 (s, 1H), 8.58 (d,1H), 7.73 (d, 2H), 7.19-7.32 (m, 4H), 6.84 (d, 1H), 6.71 (d, 1H), 6.60(t, 1H), 6.38 (t, 1H), 3.88 (s, 3H), 2.90-3.09 (m, 2H), 2.81-2.88 (m,1H), 1.97-2.06 (m, 1H).

2.2. Synthesis of Additional Compounds

The following compounds were synthesized from appropriate startingmaterials using Protocols D, E, A, B and C above or slightly modifiedversions thereof

1-(6-Methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(12)

Retention time (min)=3.113, method [7], MS (ESI) 362.1 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.41 (s, 1H), 8.42-8.47 (m, 3H), 7.69 (d, 2H),7.53 (d, 1H), 7.28 (d, 1H), 6.89 (d, 1H), 6.71 (s, 1H), 6.56 (t, 1H),6.47 (m, 1H), 3.65 (s, 3H), 3.01-3.08 (m, 1H), 2.71-2.98 (m, 1H),2.62-2.69 (m, 1H), 2.06-2.11 (m, 1H).

N-(4-chlorophenyl)-1-(6-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(13)

Retention time (min)=9.412, method [7], MS (ESI) 395.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.18 (s, 1H), 8.59 (dd, 1H), 7.75 (d, 2H), 7.19-7.32(m, 4H), 6.90 (d, 1H), 6.55-6.61 (m, 2H), 6.40 (t, 1H), 3.75 (s, 3H),2.90-3.08 (m, 2H), 2.78-2.88 (m, 1H), 1.97-2.09 (m, 1H).

N-(4-chlorophenyl)-1-(5-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(14)

Retention time (min)=9.404, method [7], MS (ESI) 417.1 (M+Na); ¹H NMR(300 MHz, CDCl₃) δ 12.19 (s, 1H), 8.58 (dd, 1H), 7.74 (d, 2H), 7.32 (d,2H), 7.18 (d, 1H), 7.06 (d, 1H), 6.89 (s, 1H), 6.83 (d, 1H), 6.49 (m,1H), 6.37 (t, 1H), 3.83 (s, 3H), 2.90-3.13 (m, 2H), 2.78-2.87 (m, 1H),1.99-2.09 (m, 1H).

1-(6-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(15)

Retention time (min)=3.067, method [7], MS (ESI) 362.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.39 (s, 1H), 8.50-8.59 (m, 3H), 7.68 (d, 2H), 7.23(d, 1H), 7.04 (d, 1H), 6.84 (s, 1H), 6.50 (d, 1H), 6.47 (m, 1H), 6.38(t, 1H), 3.82 (s, 3H), 2.91-3.11 (m, 2H), 2.79-2.88 (m, 1H), 1.99-2.10(m, 1H).

1-(6-bromo-2,3-dihydro-1H-inden-1-yl)-N-(4-chlorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide(16)

Retention time (min)=10.360, method [7], MS (ESI) 445.0 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.11 (s, 1H), 8.59 (dd, 1H), 7.73 (d, 2H), 7.46 (d,1H), 7.20-7.33 (m, 5H), 6.61 (t, 1H), 6.45 (t, 1H), 2.91-3.08 (m, 2H),2.79-2.89 (m, 1H), 2.07-2.11 (m, 1H).

1-(6-bromo-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(17)

Retention time (min)=3.842, method [7], MS (ESI) 411.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.27 (s, 1H), 8.61 (d, 1H), 8.58 (d, 2H), 7.58 (d,2H), 7.46 (d, 1H), 7.18-7.38 (m, 2H), 6.61 (t, 1H), 6.44 (t, 1H),2.90-3.14 (m, 2H), 2.80-2.89 (m, 1H), 1.84-2.09 (m, 1H).

1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(18)

Retention time (min)=3.095, method [7], MS (ESI) 362.2 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.37 (s, 1H), 8.41-8.47 (m, 3H), 7.69 (d, 2H),7.58 (d, 1H), 7.25 (t, 1H), 6.93 (d, 1H), 6.70 (d, 1H), 6.49-6.58 (m,2H), 3.81 (s, 3H), 3.00-3.08 (m, 1H), 2.71-2.98 (m, 1H), 2.65-2.69 (m,1H), 2.03-2.12 (m, 1H).

1-(7-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(19)

Retention time (min)=2.848, method [7], MS (ESI) 362.0 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.43 (s, 1H), 8.42-8.47 (m, 3H), 7.69 (d, 2H),7.00-7.39 (m, 2H), 6.97 (d, 1H), 6.88 (d, 1H), 6.51 (t, 1H), 6.38 (m,1H), 3.61 (s, 3H), 2.90-3.08 (m, 1H), 2.71-2.88 (m, 1H), 2.61-2.69 (m,1H), 1.97-2.04 (m, 1H).

N-(7-chlorophenyl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(20)

Retention time (min)=8.530, method [7], MS (ESI) 395.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.22 (s, 1H), 8.57 (d, 1H), 7.75 (d, 2H), 7.27-7.39(m, 3H), 7.10 (d, 1H), 6.97 (d, 1H), 6.74 (d, 1H), 6.49 (m, 1H), 6.38(t, 1H), 3.68 (s, 3H), 2.94-3.15 (m, 2H), 2.75-2.88 (m, 1H), 2.04-2.11(m, 1H).

N-(4-chlorophenyl)-1-(3-methyl-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(21)

Retention time (min)=9.284, method [7], MS (ESI) 379.0 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.16 (s, 1H), 8.62 (dd, 1H), 7.74 (d, 2H), 7.24-7.45(m, 6H), 6.98 (d, 1H), 6.63 (m, 1H), 6.46 (t, 1H), 3.19-3.39 (m, 1H),2.97-3.12 (m, 1H), 1.52 (m, 1H), 1.46 (d, 3H)

1-(3-methyl-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(22)

Retention time (min)=3.413, method [7], MS (ESI) 346.1 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.99 (s, 1H), 8.76 (d, 2H), 8.63 (d, 1H), 8.21 (d,2H), 7.85 (d, 1H), 7.31-7.49 (m, 3H), 7.26 (t, 1H), 6.97 (d, 1H), 6.68(t, 1H), 6.49-6.57 (m, 1H), 3.22-3.33 (m, 1H), 2.84-2.95 (m, 1H),2.65-2.69 (m, 1H), 1.65-1.78 (m, 1H), 1.36 (d, 3H).

1-(4-bromo-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(23)

Retention time (min)=3.715, method [7], MS (ESI) 411.0 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.34 (s, 1H), 8.52-8.61 (m, 3H), 7.67 (d, 2H),7.52 (d, 1H), 7.27 (m, 1H), 7.15 (t, 1H), 7.05 (d, 1H), 6.71 (t, 1H),6.45 (t, 1H), 2.91-3.22 (m, 2H), 2.84-2.90 (m, 1H), 2.00-2.11 (m, 1H).

1-(4-bromo-2,3-dihydro-1H-inden-1-yl)-N-(4-chlorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide(24)

Retention time (min)=9.577, method [7], MS (ESI) 395.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.08 (s, 1H), 8.57 (dd, 1H), 7.69 (d, 2H), 7.49 (d,1H), 7.28 (d, 2H), 7.19 (m, 1H), 7.13 (t, 1H), 7.02 (d, 1H), 6.68 (t,1H), 6.40 (t, 1H), 2.85-3.19 (m, 2H), 2.79-2.84 (m, 1H), 1.97-2.08 (m,1H).

1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(25)

The title compound (enantiomer A) was obtained through chiral separationof the racemic mixture. Retention time (min)=6.118, method [7], MS (ESI)362.1 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.37 (s, 1H), 8.58 (dd, 1H),8.52 (d, 2H), 7.69 (d, 2H), 7.23-7.29 (m, 2H), 6.85 (d, 1H), 6.71 (d,1H), 6.60 (m, 1H), 6.40 (t, 1H), 3.88 (s, 3H), 2.91-3.09 (m, 2H),2.79-2.89 (m, 1H), 1.96-2.07 (m, 1H).

1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(26)

The title compound (enantiomer B) was obtained through chiral separationof the racemic mixture. Retention time (min)=6.123, method [7], MS (ESI)362.1 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.34 (s, 1H), 8.55 (dd, 1H),8.49 (d, 2H), 7.65 (d, 2H), 7.20-7.26 (m, 2H), 6.82 (d, 1H), 6.69 (d,1H), 6.58 (m, 1H), 6.37 (t, 1H), 3.85 (s, 3H), 2.91-3.08 (m, 2H),2.76-2.88 (m, 1H), 1.93-2.04 (m, 1H).

N-(biphenyl-4-yl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(27)

Retention time (min)=10.542, method [7], MS (ESI) 437.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.19 (s, 1H), 8.59 (dd, 1H), 7.82 (d, 2H), 7.56-7.59(m, 4H), 7.30-7.42 (m, 2H), 7.17-7.28 (m, 3H), 6.82 (d, 1H), 6.70 (d,1H), 6.61 (m, 1H), 6.36 (t, 1H), 3.86 (s, 3H), 2.91-3.09 (m, 2H),2.77-2.89 (m, 1H), 1.94-2.05 (m, 1H).

1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(4-phenoxyphenyl)-1,2-dihydropyridine-3-carboxamide(28)

Retention time (min)=10.455, method [7], MS (ESI) 453.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.09 (s, 1H), 8.59 (dd, 1H), 7.76 (d, 2H), 7.18-7.32(m, 4H), 6.95-7.12 (m, 5H), 6.84 (d, 1H), 6.74 (d, 1H), 6.63 (m, 1H),6.37 (t, 1H), 3.91 (s, 3H), 2.95-3.12 (m, 2H), 2.78-2.93 (m, 1H),1.96-2.08 (m, 1H).

N-(4-bromophenyl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(29)

Retention time (min)=9.797, method [7], MS (ESI) 441.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.19 (s, 1H), 8.58 (dd, 1H), 7.68 (d, 2H), 7.45 (d,2H), 7.19-7.28 (m, 2H), 6.84 (d, 1H), 6.71 (d, 1H), 6.60 (m, 1H), 6.38(t, 1H), 3.88 (s, 3H), 2.86-3.09 (m, 2H), 2.78-2.84 (m, 1H), 1.95-2.06(m, 1H).

1-(4-isopropoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(30)

Retention time (min)=4.451, method [7], MS (ESI) 390.1 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.39 (s, 1H), 8.58 (dd, 1H), 8.52 (d, 2H), 7.68(d, 2H), 7.19-7.28 (m, 2H), 6.83 (d, 1H), 6.68 (d, 1H), 6.57 (m, 1H),6.40 (t, 1H), 4.56-4.64 (m, 2H), 2.90-3.10 (m, 2H), 2.77-2.86 (m, 1H),1.94-2.05 (m, 1H), 1.35-1.40 (m, 3H).

N-(4-chlorophenyl)-1-(4-isopropoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(31)

Retention time (min)=10.240, method [7], MS (ESI) 423.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.16 (s, 1H), 8.55 (dd, 1H), 7.70 (d, 2H), 7.16-7.30(m, 4H), 6.81 (d, 1H), 6.64 (d, 1H), 6.54 (m, 1H), 6.36 (t, 1H),4.53-4.61 (m, 2H), 2.83-3.05 (m, 2H), 2.74-2.81 (m, 1H), 1.90-2.02 (m,1H), 1.32-1.37 (m, 3H).

1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(4-phenoxyphenyl)-1,2-dihydropyridine-3-carboxamide(32)

The title compound (enantiomer A) was obtained through chiral separationof the racemic mixture. Retention time (min)=10.451, method [7], MS(ESI) 453.2 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.09 (s, 1H), 8.59 (dd,1H), 7.74 (d, 2H), 7.18-7.34 (m, 4H), 6.98-7.09 (m, 5H), 6.84 (d, 1H),6.73 (d, 1H), 6.63 (m, 1H), 6.38 (t, 1H), 3.88 (s, 3H), 2.93-3.11 (m,2H), 2.79-2.91 (m, 1H), 1.96-2.07 (m, 1H).

1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(4-phenoxyphenyl)-1,2-dihydropyridine-3-carboxamide(33)

The title compound (enantiomer B) was obtained through chiral separationof the racemic mixture. Retention time (min)=10.452, method [7], MS(ESI) 453.2 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.07 (s, 1H), 8.56 (dd,1H), 7.74 (d, 2H), 7.16-7.31 (m, 4H), 6.95-7.07 (m, 5H), 6.81 (d, 1H),6.70 (d, 1H), 6.59 (m, 1H), 6.35 (t, 1H), 3.85 (s, 3H), 2.94-3.09 (m,2H), 2.76-2.90 (m, 1H), 1.93-2.074 (m, 1H).

N-(biphenyl-4-yl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(34)

The title compound (enantiomer A) was obtained through chiral separationof the racemic mixture. Retention time (min)=10.564, method [7], MS(ESI) 437.2 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.21 (s, 1H), 8.61 (dd,1H), 7.86 (d, 2H), 7.59-7.62 (m, 4H), 7.30-7.44 (m, 2H), 7.19-7.29 (m,3H), 6.84 (d, 1H), 6.73 (d, 1H), 6.62 (m, 1H), 6.39 (t, 1H), 3.89 (s,3H), 2.90-3.12 (m, 2H), 2.80-2.89 (m, 1H), 1.97-2.08 (m, 1H).

N-(biphenyl-4-yl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(35)

The title compound (enantiomer B) was obtained through chiral separationof the racemic mixture. Retention time (min)=10.564, method [7], MS(ESI) 437.2 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.21 (s, 1H), 8.58 (dd,1H), 7.83 (d, 2H), 7.55-7.59 (m, 4H), 7.39-7.42 (m, 2H), 7.17-7.31 (m,3H), 6.82 (d, 1H), 6.71 (d, 1H), 6.61 (m, 1H), 6.36 (t, 1H), 3.86 (s,3H), 2.91-3.08 (m, 2H), 2.79-2.89 (m, 1H), 1.947-2.05 (m, 1H).

1-(4-bromo-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(36)

The title compound (enantiomer A) was obtained through chiral separationof the racemic mixture. Retention time (min)=3.960, method [7], MS (ESI)411.0 (M+H);

¹H NMR (300 MHz, DMSO-d6) δ 12.32 (s, 1H), 8.60 (dd, 1H), 8.52 (d, 2H),7.67 (d, 2H), 7.52 (d, 1H), 7.27 (m, 1H), 7.15 (t, 1H), 7.05 (d, 1H),6.71 (m, 1H), 6.45 (t, 1H), 3.01-3.22 (m, 2H), 2.83-2.94 (m, 1H),2.00-2.11 (m, 1H).

1-(4-bromo-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(37)

The title compound (enantiomer B) was obtained through chiral separationof the racemic mixture. Retention time (min)=3.933, method [7], MS (ESI)411.0 (M+H);

¹H NMR (300 MHz, DMSO-d6) δ 12.32 (s, 1H), 8.60 (dd, 1H), 8.52 (d, 2H),7.67 (d, 2H), 7.52 (d, 1H), 7.27 (m, 1H), 7.15 (t, 1H), 7.05 (d, 1H),6.71 (m, 1H), 6.45 (t, 1H), 2.98-3.18 (m, 2H), 2.80-2.92 (m, 1H),2.97-2.09 (m, 1H).

1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(4-(trifluoromethoxy)phenyl)-1,2-dihydropyridine-3-carboxamide(38)

Retention time (min)=10.216, method [7], MS (ESI) 445.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.22 (s, 1H), 8.58 (d, 1H), 7.79 (d, 2H), 7.18-7.29(m, 4H), 6.84 (d, 1H), 6.72 (d, 1H), 6.60 (m, 1H), 6.39 (t, 1H), 3.88(s, 3H), 2.93-3.11 (m, 2H), 2.79-2.91 (m, 1H), 1.96-2.07 (m, 1H).

N-cyclohexyl-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(39)

Retention time (min)=7.962, method [7], MS (ESI) 367.2 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 9.83 (m, 1H), 8.49 (d, 1H), 7.20-7.25 (m, 1H),7.10-7.13 (m, 1H), 6.82 (d, 1H), 6.68 (d, 1H), 6.58 (m, 1H), 6.29 (t,1H), 3.99-4.01 (m, 1H), 3.86 (s, 3H), 2.92-3.08 (m, 2H), 2.76-2.89 (m,1H), 1.92-2.00 (m, 3H), 1.72-1.90 (m, 3H), 1.26-1.48 (m, 5H).

N-(4-fluorophenyl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(40)

Retention time (min)=7.946, method [7], MS (ESI) 379.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.09 (s, 1H), 8.57 (dd, J=2.1 Hz, 7.2 Hz, 1H),7.68-7.78 (m, 2H), 7.18-7.30 (overlap with CDCl₃, 2H), 7.02 (m, 2H),6.82 (d, J=8.1 Hz, 1H), 6.70 (d, J=7.5 Hz, 1H), 6.58 (dd, J=5.7 Hz, 8.1Hz, 1H), 6.40 (t, J=7.2 Hz, 1H), 3.87 (s, 3H), 2.75-3.10 (m, 3H),1.95-2.05 (m, 1H).

N-(4-iodophenyl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(41)

Retention time (min)=9.596, method [7], MS (ESI) 487.0 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.10 (s, 1H), 8.60 (dd, J=2.1 Hz, 7.2 Hz, 1H), 7.67(d, J=8.7 Hz, 2H), 7.59 (d, J=8.7 Hz, 2H), 7.20-7.35 (overlap withCDCl₃, 2H), 6.87 (d, J=7.8 Hz, 1H), 6.74 (d, J=7.8 Hz, 1H), 6.60 (dd,J=5.7 Hz, 7.8 Hz, 1H), 6.41 (t, J=7.2 Hz, 1H), 3.92 (s, 3H), 2.80-3.14(m, 3H), 1.95-2.05 (m, 1H).

1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-phenyl-1,2-dihydropyridine-3-carboxamide(42)

Retention time (min)=8.181, method [7], MS (ESI) 361.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.05 (s, 1H), 8.60 (dq, J=7.0 Hz, 1.2 Hz, 1H), 7.67(d, J=8.4 Hz, 2H), 7.36 (t, J=8.4 Hz, 2H), 7.27 (overlap with CDCl₃,1H), 7.20 ((dq, J=7.0 Hz, 1.2 Hz, 1H), 7.13 (t, J=7.5 Hz, 1H), 6.85 (d,J=8.1 Hz, 1H), 6.73 (d, J=7.5 Hz, 1H), 6.63 (dd, J=6.0 Hz, 7.8 Hz, 1H),6.40 (t, J=7.0 Hz, 1H), 3.90 (s, 3H), 2.78-3.16 (m, 3H), 1.95-2.05 (m,1H).

N-(4,4-difluorocyclohexyl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(43)

Retention time (min)=7.374, method [7], MS (ESI) 403.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 10.00 (d, J=7.2 Hz, 1H), 8.49 (dd, J=7.0 Hz, 2.4 Hz,1H), 7.26 (overlap with CDCl₃, 1H), 7.16 ((dd, J=7.0, 2.4 Hz, 1H), 6.83(d, J=8.1 Hz, 1H), 6.69 (d, J=7.5 Hz, 1H), 6.56 (dd, J=5.7 Hz, 8.1 Hz,1H), 6.33 (t, J=7.0 Hz, 1H), 4.05-4.20 (m, 1H), 3.88 (s, 3H), 2.78-3.16(m, 3H), 1.85-2.12 (m, 9H).

N-(3,4-difluorophenyl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(44)

Retention time (min)=9.140, method [7], MS (ESI) 419.1 (M+Na); ¹H NMR(300 MHz, CDCl₃) δ 12.22 (s, 1H), 8.58 (dd, J=2.1 Hz, 7.0 Hz, 1H),7.87-7.95 (m, 1H), 7.20-7.35 (overlap with CDCl₃, 2H), 7.13 (q, J=8.7Hz, 1H), 7.02 (m, 2H), 6.86 (d, J=8.1 Hz, 1H), 6.73 (d, J=7.8 Hz, 1H),6.60 (dd, J=5.4 Hz, 8.4 Hz, 1H), 6.41 (t, J=7.0 Hz, 1H), 3.89 (s, 3H),2.78-3.13 (m, 3H), 1.95-2.05 (m, 1H).

1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-N-(4-(4-methoxyphenoxy)phenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide(45)

Retention time (min)=10.129, method [7], MS (ESI) 483.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.06 (s, 1H), 8.59 (m, 1H), 7.71 (d, J=8.7 Hz, 2H),7.20-7.35 (overlap with CDCl₃, 3H), 6.87 (t, J=9.3 Hz, 4H), 6.80-6.92(m, 3H), 6.73 (m, 1H), 6.41 (t, J=7.0 Hz, 1H), 3.89 (s, 3H), 3.80 (s,3H), 2.78-3.13 (m, 3H), 1.95-2.05 (m, 1H).

Example 3 3.1. Synthesis of1-(4-cyclopropyl-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(46) Protocol F

To a solution of 4-bromo-2,3-dihydro-1H-inden-1-one (2.2 g, 1.0 eq.) in60 ml MeOH was added NH₂OCH₃.HCl (0.914 g, 1.15 eq.) and sodium acetate(0.924 g, 1.18 eq). The resulting mixture was stirred at roomtemperature overnight. The reaction solvent was removed in vacuo. Theresidue was diluted with 20 ml water and extracted by 2×50 ml ethylacetate. The combined organic phase was washed with water, brine anddried over sodium sulfate. The solvent was removed in vacuo to affordclear oil 2.45 g (yield 98%) 4-bromo-2,3-dihydro-1H-inden-1-one O-methyloxime which was used to next step without further purification.

4-bromo-2,3-dihydro-1H-inden-1-one O-methyl oxime (1.0 g, 1.0 eq.),cyclopropylboronic acid (0.537, 1.5 eq.), Palladium acetate (0.094 g,0.1 eq.), K₃PO₄ (3.09 g, 3.5 eq.) and tricyclohexylphosphine (0.234 g,0.2 eq.) were combined in 50 ml Toluene and 1.6 ml water. The reactionmixture was heated at 100° C. for 3 hrs. The reaction mixture wasfiltered through a celite pad to remove inorganic salt. The filtrate wasconcentrated in vacuo, and the residue was directly purified on flashcolumn to provide yellow oil 0.828 g (yield 99%)4-cyclopropyl-2,3-dihydro-1H-inden-1-one O-methyl oxime.

Protocol H

A solution of 4-cyclopropyl-2,3-dihydro-1H-inden-1-one O-methyl oxime(0.828 g) in 50 ml MeOH was saturated with NH₃ gas at 0° C., then added3 ml Raney-Nickel in water. The mixture was hydrogenated (60 psi H2) atambient temperature for 3 hrs. The catalyst was removed by filtration.The solvent was concentrated in vacuo to give green oil 0.6 g (yield84%) 4-cyclopropyl-2,3-dihydro-1H-inden-1-amine which was used to nextstep without further purification.

Following Protocol A, B andC₁₋₄-cyclopropyl-2,3-dihydro-1H-inden-1-amine was converted to1-(4-cyclopropyl-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide.Retention time (min)=4.435, method [7],

MS (ESI) 372.2 (M+H); ¹H NMR (300 MHz, DMSO-d6) δ 12.41 (s, 1H),8.42-8.47 (m, 3H), 7.69 (d, 2H), 7.59 (d, 1H), 4.14 (t, 1H), 6.88 (d,1H), 6.84 (d, 1H), 6.51-6.59 (m, 2H), 3.19 (m, 1H), 3.05 (m, 1H), 2.68(m, 1H), 2.10 (m, 1H), 1.90 (m, 1H), 0.90-0.97 (m, 2H), 0.63-0.75 (m,2H).

3.2. Synthesis ofN-(4-chlorophenyl)-1-(4-cyclopropyl-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(47)

The title compound was synthesized from appropriate starting materialsusing Protocols F, H, A, B and C above or slightly modified versionsthereof.

Retention time (min)=10.978, method [7], MS (ESI) 405.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.13 (s, 1H), 8.59 (dd, 1H), 7.74 (d, 2H), 7.30 (d,1H), 7.17-7.23 (m, 2H), 6.86-6.93 (m, 2H), 6.61 (m, 1H), 6.39 (t, 1H),3.05-3.23 (m, 2H), 2.82-2.94 (m, 1H), 1.98-2.10 (m, 1H), 1.86-1.95 (m,1H), 0.97-1.24 (m, 1H), 0.68-0.79 (m, 2H).

Example 4 4.1. Synthesis of1-(7,8-dihydro-6H-indeno[5,4-d][1,3]dioxol-6-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(48)

Benzo[d][1,3]dioxole-4-carbaldehyde (5.0 g, 1.0 eq.) was dissolved in 15ml pyridine, then added malonic acid (6.58 g, 1.9 eq.) and piperidine(0.284 g, 0.1 eq.). The resulting mixture was heated at 105° C. for 3hrs. After cooling to room temperature, the reaction mixture was pouredinto a solution of 25 ml concentrated HCl in 300 g ice. The whiteprecipitate was collected by filtration; the solid was washed with 3×30ml water and dried in air to afford white solid 6.2 g (yield 97%)(E)-3-(benzo[d][1,3]dioxol-4-yl)acrylic acid which was used withoutfurther purification.

(E)-3-(benzo[d][1,3]dioxol-4-yl)acrylic acid was dissolved (5.5 g) in150 ml 95% EtOH, added catalyst (0.55 g, 10% Palladium on carbon). Themixture was hydrogenated (60 psi H2) at ambient temperature for 3 hrs.The catalyst was removed by filtration. The solvent was evaporated invacuo to give white solid 5 g (yield 90%)3-(benzo[d][1,3]dioxol-4-yl)propanoic acid which was used withoutfurther purification.

To a vigorously stirred mixture of P₂O₅ (15.6 g) in 100 ml dry benzeneat reflux was added a solution of 3-(benzo[d][1,3]dioxol-4-yl)propanoicacid (5 g) in 50 ml benzene dropwise. The reaction kept refluxing for 45mins, additional P₂O₅ (10 g) was added. After one hour, the reactionmixture was poured into 200 ml ice-water. The organic phase wasseparated and washed with 50 ml 1N NaOH aqueous solution and brine. Thesolvent was evaporated in vacuo to afford dark oil. The oil was purifiedon flash column in silica gel to provide yellow solid 0.889 g (yield20%) 7,8-dihydro-6H-indeno[5,4-d][1,3]dioxol-6-one.

Following Protocol F, H, A, B and C,7,8-dihydro-6H-indeno[5,4-d][1,3]dioxol-6-one was converted to1-(7,8-dihydro-6H-indeno[5,4-d][1,3]dioxol-6-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide.Retention time (min)=2.895, method [7], MS (ESI) 376.1 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.39 (s, 1H), 8.41-8.47 (m, 3H), 7.68 (d, 2H),7.60 (d, 1H), 6.82 (d, 1H), 6.65 (d, 1H), 6.53 (t, 1H), 6.38-6.42 (m,1H), 6.05 (s, 3H), 2.91-3.08 (m, 1H), 2.83-2.89 (m, 1H), 2.63-2.675 (m,1H), 2.02-2.17 (m, 1H).

4.2. Additional Compounds

The following compounds were synthesized from appropriate startingmaterials using the procedures outlined above in Example 4.1., orslightly modified versions thereof.

N-(4-chlorophenyl)-1-(7,8-dihydro-6H-indeno[5,4-d][1,3]dioxol-6-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(49)

Retention time (min)=9.105, method [7], MS (ESI) 431.1 (M+Na); ¹H NMR(300 MHz, CDCl₃) δ 12.15 (s, 1H), 8.58 (dd, 1H), 7.73 (d, 2H), 7.31 (d,2H), 7.20-7.28 (m, 1H), 6.77 (d, 1H), 6.47 (m, 1H), 6.39 (t, 1H), 6.02(s, 2H), 2.97-3.03 (m, 2H), 2.78-2.90 (m, 1H), 2.01-2.14 (m, 1H).

N-(4-chlorophenyl)-2-oxo-1-(4-(trifluoromethoxy)-2,3-dihydro-1H-inden-1-yl)-1,2-dihydropyridine-3-carboxamide(50)

Retention time (min)=10.805, method [7], MS (ESI) 449.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.15 (s, 1H), 8.60 (d, 1H), 7.72 (d, 2H), 7.21-7.35(m, 5H), 7.05 (d, 1H), 6.68 (t, 1H), 6.44 (t, 1H), 3.03-3.26 (m, 2H),2.84-2.96 (m, 1H), 2.02-2.14 (m, 1H).

2-oxo-N-(pyridin-4-yl)-1-(4-(trifluoromethoxy)-2,3-dihydro-1H-inden-1-yl)-1,2-dihydropyridine-3-carboxamide(51)

Retention time (min)=4.679, method [7], MS (ESI) 416.1 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.89 (s, 1H), 8.70 (d, 2H), 8.49 (dd, 1H), 8.17(d, 2H), 7.82 (d, 1H), 7.31-7.41 (m, 2H), 7.17 (d, 1H), 6.55-6.66 (m,2H), 3.16-3.26 (m, 1H), 2.96-3.07 (m, 1H), 2.68-2.80 (m, 1H), 2.14-2.26(m, 1H).

Example 5 Synthesis of1-(4-(methylthio)-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(52)

Following Protocol F, 4-fluoro-2,3-dihydro-1H-inden-1-one was convertedto 4-fluoro-2,3-dihydro-1H-inden-1-one O-methyl oxime.

4-fluoro-2,3-dihydro-1H-inden-1-one O-methyl oxime (0.5 g, 1.0 eq) andsodium methanethiolate (0.215 g, 1.1 eq.) were combined in 20 ml dryDMF. The mixture was heated at 130° C. for 7 hrs under N₂ atmosphere.After reaction cooled to room temperature, the mixture was diluted with60 ml ethyl acetate and 60 ml hexane, and washed with 3×50 ml water andbrine. The crude mixture was purified on flash column in silica gel toprovide white solid 0.47 g (yield 81%)4-(methylthio)-2,3-dihydro-1H-inden-1-one O-methyl oxime.

A solution of 4-(methylthio)-2,3-dihydro-1H-inden-1-one O-methyl oxime(0.47 g, 1.0 eq.) in 10 ml dry THF was cooled to 0° C. under N₂atmosphere. Then Borane/THF complex (15 ml, 1M, 6.6 eq.) was added toreaction dropwise. The resulting mixture was stirred at room temperaturefor 10 minutes, and heated at refluxing overnight. The reaction wasquenched by adding 100 ml ice-water. Then the mixture was extracted with2×50 ml ethyl acetate. The combined organic phase was washed with 2NNa₂CO₃, and brine. Removal of solvent in vacuo afforded an oil 0.22 g(yield 54%) 4-(methylthio)-2,3-dihydro-1H-inden-1-amine which was usedwithout further purification.

Following Protocol A, B and C,4-(methylthio)-2,3-dihydro-1H-inden-1-amine was converted to1-(4-(methylthio)-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide.Retention time (min)=3.590, method [7],

MS (ESI) 378.1 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.35 (s, 1H), 8.59 (dd,1H), 8.52 (d, 2H), 7.69 (d, 2H), 7.23-7.31 (m, 2H), 7.16 (d, 1H), 6.89(d, 1H), 6.62 (m, 1H), 6.42 (t, 1H), 3.00-3.08 (m, 1H), 2.92-3.08 (m,1H), 2.84-2.91 (m, 1H), 2.53 (s, 3H), 1.99-2.11 (m, 1H).

Example 6 6.1. Synthesis of tert-butyl4-(1-(2-oxo-3-(pyridin-4-ylcarbamoyl)pyridin-1(2H)-yl)-2,3-dihydro-1H-inden-4-yl)piperazine-1-carboxylate(53)

4-bromo-2,3-dihydro-1H-inden-1-one O-methyl oxime (2.5 g, 1.0 eq.),tert-butyl piperazine-1-carboxylate (2.91 g, 1.5 eq.), Pd₂(dba)₃ (0.76g, 0.08 eq.), BINAP (0.63 g, 0.1 eq.) and Sodium t-butoxide (1.6 g, 1.6eq.) were combined in 30 ml 1,4-dioxane. The reaction mixture was heatedat 100° C. for 2.5 hrs under N₂. The reaction mixture was filteredthrough a celite pad to remove inorganic salt. The filtrate wasconcentrated in vacuo, and the residue mixture was directly purified onflash column to provide yellow solid 2.7 g (yield 75%) tert-butyl4-(1-(methoxyimino)-2,3-dihydro-1H-inden-4-yl)piperazine-1-carboxylate.

Following Protocol H, tert-butyl4-(1-(methoxyimino)-2,3-dihydro-1H-inden-4-yl)piperazine-1-carboxylatewas converted to tert-butyl4-(1-amino-2,3-dihydro-1H-inden-4-yl)piperazine-1-carboxylate.

Following Protocol A, B and C, to tert-butyl4-(1-amino-2,3-dihydro-1H-inden-4-yl)piperazine-1-carboxylate wasconverted to tert-butyl4-(1-(2-oxo-3-(pyridin-4-ylcarbamoyl)pyridin-1(2H)-yl)-2,3-dihydro-1H-inden-4-yl)piperazine-1-carboxylate( ) Retention time (min)=5.050, method [7], MS (ESI) 516.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.34 (s, 1H), 8.57 (dd, 1H), 8.50 (d, 2H), 7.66 (d,2H), 7.20-7.27 (m, 2H), 6.87 (d, 1H), 6.74 (d, 1H), 6.56 (m, 1H), 6.39(t, 1H), 3.54-3.57 (m, 4H), 2.91-3.08 (m, 6H), 2.76-2.88 (m, 1H),1.95-2.06 (m, 1H), 1.46 (s, 9H).

6.2. Additional Compounds

The following compounds were synthesized from appropriate startingmaterials using the procedures outlined above in Example 6.1., orslightly modified versions thereof.

tert-butyl4-(1-(3-(4-chlorophenylcarbamoyl)-2-oxopyridin-1(2H)-yl)-2,3-dihydro-1H-inden-4-yl)piperazine-1-carboxylate(54)

Retention time (min)=10.340, method [7], MS (ESI) 549.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.15 (s, 1H), 8.59 (dd, 1H), 7.73 (d, 2H), 7.32 (d,2H), 7.20-7.28 (m, 2H), 6.89 (d, 1H), 6.76 (d, 1H), 6.60 (m, 1H), 6.40(t, 1H), 3.57-3.60 (m, 4H), 2.90-3.09 (m, 6H), 2.78-2.89 (m, 1H),1.97-2.09 (m, 1H), 1.48 (s, 9H).

1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(55)

Retention time (min)=2.219, method [7], MS (ESI) 417.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.37 (s, 1H), 8.59 (dd, 1H), 8.52 (d, 2H), 7.68 (d,2H), 7.23-7.29 (m, 2H), 6.92 (d, 1H), 6.76 (d, 1H), 6.58 (t, 1H), 6.42(t, 1H), 3.83-3.88 (m, 4H), 2.87-3.13 (m, 6H), 2.78-2.86 (m, 1H),2.00-2.09 (m, 1H).

N-(4-chlorophenyl)-1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(56)

Retention time (min)=8.265, method [7], MS (ESI) 450.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.13 (s, 1H), 8.56 (dd, 1H), 7.70 (d, 2H), 7.20-7.30(m, 4H), 6.88 (d, 1H), 6.73 (d, 1H), 6.56 (t, 1H), 6.37 (t, 1H),3.80-3.85 (m, 4H), 2.83-3.10 (m, 6H), 2.74-2.81 (m, 1H), 1.93-2.06 (m,1H).

1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(4-phenoxyphenyl)-1,2-dihydropyridine-3-carboxamide(57)

Retention time (min)=9.261, method [7], MS (ESI) 508.3 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.08 (s, 1H), 8.57 (dd, 1H), 7.72 (d, 2H), 7.19-7.31(m, 4H), 6.95-7.06 (m, 5H), 6.88 (d, 1H), 6.74 (d, 1H), 6.56 (t, 1H),6.37 (t, 1H), 3.80-3.85 (m, 4H), 2.92-3.10 (m, 6H), 2.75-2.89 (m, 1H),1.95-2.13 (m, 1H).

N-(4-fluorophenyl)-1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(58)

Retention time (min)=7.109, method [7], MS (ESI) 434.2 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.11 (s, 1H), 8.43 (dd, 1H), 7.71-7.75 (m, 2H),7.57 (m, 1H), 7.15-7.21 (m, 3H), 6.88 (d, 1H), 6.71 (d, 1H), 6.49-6.57(m, 2H), 3.68-3.77 (m, 4H), 2.85-3.07 (m, 6H), 2.60-2.71 (m, 1H),2.02-2.13 (m, 1H).

N-(4-bromophenyl)-1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(59)

Retention time (min)=8.050, method [7], MS (ESI) 495.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.16 (s, 1H), 8.56 (dd, 1H), 7.66 (d, 2H), 7.43 (d,2H), 7.20-7.25 (m, 2H), 6.88 (d, 1H), 6.73 (d, 1H), 6.56 (t, 1H), 6.37(t, 1H), 3.82-3.85 (m, 4H), 2.90-3.10 (m, 6H), 2.77-2.87 (m, 1H),2.00-2.06 (m, 1H).

N-cyclohexyl-1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(60)

Retention time (min)=6.326, method [7], MS (ESI) 422.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 9.79 (d, 1H), 8.48 (dd, 1H), 7.10-7.20 (m, 2H), 6.86(d, 1H), 6.70 (d, 1H), 6.54 (t, 1H), 6.28 (t, 1H), 3.95-4.04 (m, 1H),3.81-3.85 (m, 4H), 2.73-3.09 (m, 6H), 1.90-2.02 (m, 3H), 1.71-1.78 (m,2H), 1.60-1.69 (m, 1H), 1.21-1.46 (m, 5H).

N-(4-iodophenyl)-1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(61)

Retention time (min)=8.637, method [7], MS (ESI) 542.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.17 (s, 1H), 8.59 (dd, 1H), 7.64 (d, 2H), 7.56 (d,2H), 7.23-7.28 (m, 2H), 6.91 (d, 1H), 6.75 (d, 1H), 6.59 (t, 1H), 6.39(t, 1H), 3.85-3.88 (m, 4H), 2.88-3.13 (m, 6H), 2.77-2.86 (m, 1H),1.97-2.09 (m, 1H).

1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-p-tolyl-1,2-dihydropyridine-3-carboxamide(62)

Retention time (min)=7.330, method [7], MS (ESI) 430.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.00 (s, 1H), 8.57 (dd, 1H), 7.63 (d, 2H), 7.18-7.25(m, 2H), 7.13 (d, 2H), 6.88 (d, 1H), 6.73 (d, 1H), 6.58 (t, 1H), 6.35(t, 1H), 3.82-3.85 (m, 4H), 2.92-3.10 (m, 6H), 2.76-2.89 (m, 1H), 2.30(s, 3H), 1.95-2.06 (m, 1H).

1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-phenyl-1,2-dihydropyridine-3-carboxamide(63)

Retention time (min)=6.803, method [7], MS (ESI) 416.3 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.09 (s, 1H), 8.64 (dd, 1H), 7.79-7.82 (m, 2H),7.31-7.40 (m, 2H), 7.24-7.28 (m, 2H), 7.16 (m, 1H), 6.93 (d, 1H), 6.80(d, 1H), 6.64 (t, 1H), 6.42 (t, 1H), 3.86-3.91 (m, 4H), 2.98-3.16 (m,6H), 2.81-2.95 (m, 1H), 2.01-2.12 (m, 1H).

1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-phenyl-1,2-dihydropyridine-3-carboxamide(64)

Retention time (min)=9.430, method [7], MS (ESI) 492.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.16 (s, 1H), 8.62 (dd, 1H), 7.85 (d, 2H), 7.58-7.61(m, 4H), 7.40-7.45 (m, 2H), 7.23-7.33 (m, 3H), 6.91 (d, 1H), 6.77 (d,1H), 6.62 (t, 1H), 6.40 (t, 1H), 3.83-3.89 (m, 4H), 2.90-3.13 (m, 6H),2.79-2.89 (m, 1H), 1.99-2.11 (m, 1H).

N-(4,4-difluorocyclohexyl)-1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(65)

Retention time (min)=6.038, method [7], MS (ESI) 458.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 9.98 (d, 1H), 8.47 (dd, 1H), 7.14-7.21 (m, 2H), 6.86(d, 1H), 6.70 (d, 1H), 6.52 (t, 1H), 6.30 (t, 1H), 4.07-4.11 (m, 1H),3.82-3.85 (m, 4H), 2.90-3.09 (m, 6H), 2.74-2.87 (m, 1H), 1.66-2.11 (m,9H).

N-(4-methoxyphenyl)-1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(66)

Retention time (min)=6.165, method [7], MS (ESI) 446.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 11.99 (s, 1H), 8.63 (dd, 1H), 7.73 (d, 2H), 7.23-7.31(m, 2H), 6.90-6.95 (m, 3H), 6.79 (d, 1H), 6.63 (t, 1H), 6.341 (t, 1H),3.88-3.93 (m, 4H), 3.83 (s, 3H), 2.93-3.16 (m, 6H), 2.80-2.91 (m, 1H),2.01-2.12 (m, 1H).

N-(4-(difluoromethoxy)phenyl)-1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(67)

Retention time (min)=7.400, method [7], MS (ESI) 482.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.13 (s, 1H), 8.63 (dd, 1H), 7.79 (d, 2H), 7.23-7.31(m, 2H), 7.13 (d, 2H), 6.94 (d, 1H), 6.79 (d, 1H), 6.62 (t, 1H), 6.50(t, 1H), 6.43 (t, 1H), 3.88-3.91 (m, 4H), 2.93-3.16 (m, 6H), 2.80-2.91(m, 1H), 2.01-2.12 (m, 1H).

1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(4-(trifluoromethoxy)phenyl)-1,2-dihydropyridine-3-carboxamide(68)

Retention time (min)=8.696, method [7], MS (ESI) 500.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.21 (s, 1H), 8.60 (dd, 1H), 7.80 (d, 2H), 7.18-7.28(m, 4H), 6.92 (d, 1H), 6.76 (d, 1H), 6.59 (t, 1H), 6.40 (t, 1H),3.83-3.88 (m, 4H), 2.90-3.13 (m, 6H), 2.78-2.89 (m, 1H), 1.98-2.10 (m,1H).

1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(4-(trifluoromethyl)phenyl)-1,2-dihydropyridine-3-carboxamide(69)

Retention time (min)=8.662, method [7], MS (ESI) 484.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.35 (s, 1H), 8.58 (dd, 1H), 7.86 (d, 2H), 7.58 (d,2H), 7.21-7.26 (m, 2H), 6.89 (d, 1H), 6.73 (d, 1H), 6.57 (t, 1H), 6.38(t, 1H), 3.80-3.86 (m, 4H), 2.90-3.10 (m, 6H), 2.75-2.88 (m, 1H),1.96-2.07 (m, 1H).

N-(4-cyanophenyl)-1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(70)

Retention time (min)=6.687, method [7], MS (ESI) 441.2 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.49 (s, 1H), 8.44 (dd, 1H), 7.89 (d, 2H), 7.80(d, 2H), 7.61-7.63 (m, 1H), 7.19 (t, 1H), 6.88 (d, 1H), 6.72 (d, 1H),6.49-6.59 (m, 2H), 3.68-3.77 (m, 4H), 2.85-3.07 (m, 6H), 2.58-2.70 (m,1H), 2.04-2.14 (m, 1H).

1-(4-(4-hydroxypiperidin-1-yl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(71)

MS (ESI) 431.2 (M+H); ¹H NMR (300 MHz, DMSO-d6) δ 12.38 (s, 1H),8.42-8.47 (m, 3H), 7.61-7.69 (m, 3H), 7.14 (t, 1H), 6.86 (d, 1H), 6.66(d, 1H), 6.48-6.59 (m, 2H), 4.68 (d, 1H), 3.57-3.64 (m, 1H), 3.12-3.23(m, 2H), 2.88-3.06 (m, 2H), 2.46-2.83 (m, 3H), 2.03-2.12 (m, 1H),1.76-1.84 (m, 2H), 1.46-1.57 (m, 2H).

N-(4-chlorophenyl)-1-(4-(4-hydroxypiperidin-1-yl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(72)

Retention time (min)=4.433, method [7], MS (ESI) 464.2 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.23 (s, 1H), 8.42 (dd, 1H), 7.75 (d, 2H),7.58-7.61 (m, 1H), 7.38 (d, 2H), 7.14 (t, 1H), 6.86 (d, 1H), 6.65 (d,1H), 6.48-6.57 (m, 2H), 4.68 (d, 1H), 3.57-3.64 (m, 1H), 3.15-3.23 (m,2H), 2.93-3.06 (m, 2H), 2.46-2.91 (m, 3H), 2.03-2.12 (m, 1H), 1.81-1.86(m, 2H), 1.46-1.56 (m, 2H).

N-(4-(4-methoxyphenylamino)phenyl)-1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(73)

Retention time (min)=7.749, method [7], MS (ESI) 537.2 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 11.96 (s, 1H), 8.40 (dd, 1H), 7.81 (s, 1H),7.50-7.56 (m, 3H), 7.18 (t, 1H), 7.00 (d, 2H), 6.82-6.92 (m, 5H), 6.70(d, 1H), 6.49-6.55 (m, 2H), 3.71-3.74 (m, 4H), 3.68 (s, 3H), 2.85-3.08(m, 6H), 2.58-2.69 (m, 1H), 2.01-2.13 (m, 1H).

N-(3,4-difluorophenyl)-1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(74)

Retention time (min)=7.787, method [7], MS (ESI) 452.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.20 (s, 1H), 8.59 (dd, 1H), 7.85-7.93 (m, 1H),7.24-7.33 (m, 3H), 7.06-7.16 (m, 1H), 6.92 (d, 1H), 6.75 (d, 1H), 6.58(t, 1H), 6.41 (t, 1H), 3.85-3.88 (m, 4H), 2.86-3.13 (m, 6H), 2.77-2.84(m, 1H), 1.98-2.09 (m, 1H).

tert-butyl4-(1-(3-(4-fluorophenylcarbamoyl)-2-oxopyridin-1(2H)-yl)-2,3-dihydro-1H-inden-4-yl)piperazine-1-carboxylate(75)

Retention time (min)=9.762, method [7], MS (ESI) 533.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.06 (s, 1H), 8.57 (dd, 1H), 7.68-7.72 (m, 2H),7.19-7.24 (m, 2H), 6.98-7.04 (m, 2H), 6.86 (d, 1H), 6.73 (d, 1H), 6.57(t, 1H), 6.37 (t, 1H), 3.53-3.59 (m, 4H), 2.91-3.08 (m, 6H), 2.78-2.89(m, 1H), 1.99-2.06 (m, 1H), 1.45 (s, 9H).

tert-butyl4-(1-(3-(4-chlorophenylcarbamoyl)-2-oxopyridin-1(2H)-yl)-2,3-dihydro-1H-inden-4-yl)piperazine-1-carboxylate(76)

Retention time (min)=11.439, method [7], MS (ESI) 594.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.14 (s, 1H), 8.56 (dd, 1H), 7.65 (d, 2H), 7.42 (d,2H), 7.19-7.24 (m, 2H), 6.86 (d, 1H), 6.73 (d, 1H), 6.54 (t, 1H), 6.37(t, 1H), 3.54-3.57 (m, 4H), 2.87-3.08 (m, 6H), 2.75-2.86 (m, 1H),1.94-2.06 (m, 1H), 1.45 (s, 9H).

N-(4-(dimethylamino)phenyl)-1-(4-morpholino-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(77)

Retention time (min)=3.575, method [7], MS (ESI) 459.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 11.87 (s, 1H), 8.59 (dd, 1H), 7.65 (d, 2H), 7.18-7.27(m, 2H), 7.13 (d, 2H), 6.90 (d, 1H), 6.72-6.78 (m, 3H), 6.60 (t, 1H),6.37 (t, 1H), 3.82-3.85 (m, 4H), 2.95-3.13 (m, 6H), 2.94 (s, 6H),2.79-2.84 (m, 1H), 1.98-2.07 (m, 1H).

Example 7 Synthesis of1-(4-(cyclopropylamino)-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(78)

To a solution of1-(4-bromo-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxylicacid (0.6 g, 1.0 eq.) in 20 ml DMF was added K₂CO₃ (0.744, 3.0 eq) andiodoethane (0.56 g, 2.0 eq.) The resulting mixture was stirred at roomtemperature overnight. The reaction was diluted with 60 ml ethyl acetateand 20 ml hexane; then washed with 3×50 ml water and 50 ml brine. Thesolvent was evaporated in vacuo to give oil 0.5 g (yield 77%) ethyl1-(4-bromo-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxylatewhich was used without further purification.

Ethyl1-(4-bromo-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxylate(0.5 g, 1.0 eq.), cyclopropanamine (0.236 g, 3.0 eq.), Palladium acetate(0.062 g, 0.2 eq.), cesium carbonate (1.124 g, 2.5 eq.) and dppf (0.25g, 0.3 eq.) were combined 20 ml Toluene in a sealed-tube. The mixturewas flashed with N₂ for a minute. Then the resulting mixture was heatedat 90° C. for 5 hrs. Removal in-organic salt through filtration affordedbrown oil. The crude mixture was purified on flash column for give anoil 50 mg (yield 10%) ethyl1-(4-(cyclopropylamino)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxylate.

To a solution of Ethyl1-(4-(cyclopropylamino)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxylate(25 mg, 1.0 eq) in 2 ml MeOH and 1 ml water was added KOH (12.43 mg, 3.0eq.). The mixture was heated at 60° C. for 15 mins. The solvent wasremoved in vacuo, the residue aqueous solution was acidified to pH 2-3by adding 2N HCl. The white precipitate was collected by filtration, anddried in vacuo to provide solid 19 mg (yield 83%)1-(4-(cyclopropylamino)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxylicacid which was used without further purification.

Following Protocol C,1-(4-(cyclopropylamino)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxylicacid was converted to1-(4-(cyclopropylamino)-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide( ) Retention time (min)=3.656, method [7], MS (ESI) 387.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.36 (s, 1H), 8.48-8.56 (m, 3H), 7.21 (d, 2H),7.00-7.25 (m, 1H), 6.98 (d, 1H), 6.48-6.52 (m, 2H), 6.36 (t, 1H), 4.07(d, 1H), 2.67-2.90 (m, 2H), 2.43-2.50 (m, 1H), 1.93-2.04 (m, 1H),0.77-0.78 (m, 2H), 0.42-0.44 (m, 2H).

Example 8 8.1. Synthesis ofN-(4-chlorophenyl)-1-(4-(methylsulfonyl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(79)

1-(4-(methylthio)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxylicacid (80 mg, 1.0 eq.) was dissolved in 2 ml CH₂Cl₂. To the solution wasadded m-CPBA (115 mg, 2.5 eq.). The resulting mixture was stirred atroom temperature overnight. The crude mixture was directly purified onflash column to afford white solid 17.8 mg (yield 20%)1-(4-(methylsulfonyl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxylicacid.

Following Protocol C,1-(4-(methylsulfonyl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxylicacid was converted toN-(4-chlorophenyl)-1-(4-(methylsulfonyl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide( ) Retention time (min)=7.196, method [7], MS (ESI) 443.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.03 (s, 1H), 8.59 (dd, 1H), 7.96 (d, 1H), 7.68 (d,2H), 7.47 (t, 1H), 7.36 (d, 1H), 7.25 (d, 2H), 7.17 (d, 1H), 6.69 (t,1H), 6.43 (t, 1H), 3.58-3.68 (m, 1H), 2.25-3.36 (m, 1H), 3.08 (s, 3H),2.86-2.97 (m, 1H), 2.08-2.20 (m, 1H).

8.2. Synthesis of1-(4-(methylsulfonyl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(80)

The title compound was synthesized from appropriate starting materialsusing the procedures outlined above in Example 8.1., or slightlymodified versions thereof.

Retention time (min)=1.332, method [7], MS (ESI) 410.0 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.97 (s, 1H), 8.74 (d, 2H), 8.49 (d, 1H), 8.23 (d,2H), 7.83-7.85 (m, 2H), 7.48-7.55 (m, 2H), 6.64 (t, 1H), 6.56 (t, 1H),3.48-3.58 (m, 1H), 3.26-3.37 (m, 1H), 3.23 (s, 3H), 2.70-2.82 (m, 1H),2.17-2.29 (m, 1H).

Example 9 9.1. Synthesis ofN-(4-chlorophenyl)-2-oxo-1-(4-(piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-1,2-dihydropyridine-3-carboxamide(81)

To a solution of tert-butyl4-(1-(3-(4-chlorophenylcarbamoyl)-2-oxopyridin-1(2H)-yl)-2,3-dihydro-1H-inden-4-yl)piperazine-1-carboxylate(460 mg) in 5 ml CH₂Cl₂ was added 2 ml TFA. The mixture was stirred atroom temperature for 2 hrs. The solvent and TFA were removed in vacuo.The residue was dissolved in 50 ml ethyl acetate, washed with 2×50 m12 MNa₂CO₃ and brine. The organic phase was dried over sodium sulfate; thenevaporated in vacuo to provide white solid 338 mg (yield 90%)N-(4-chlorophenyl)-2-oxo-1-(4-(piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-1,2-dihydropyridine-3-carboxamide.Retention time (min)=5.060, method [7], MS (ESI) 449.1 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.23 (s, 1H), 8.42 (dd, 1H), 7.73 (d, 2H), 7.60(m, 1H), 7.40 (d, 2H), 7.16 (t, 1H), 6.84 (d, 1H), 6.66 (d, 1H),6.48-6.57 (m, 2H), 2.82-3.04 (m, 9H), 2.60-2.68 (m, 1H), 2.15-2.32 (m,1H), 2.03-2.12 (m, 1H).

9.2. Additional Compounds

The following compounds were synthesized from appropriate startingmaterials using the procedures outlined above in Example 9.1., orslightly modified versions thereof.

2-oxo-1-(4-(piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(82)

Retention time (min)=8.285, method [6], MS (ESI) 416.2 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.37 (s, 1H), 8.42-8.47 (m, 3H), 7.66 (d, 2H),7.62 (m, 1H), 7.16 (t, 1H), 6.85 (d, 1H), 6.68 (d, 1H), 6.48-6.57 (m,2H), 2.83-3.06 (m, 10H), 2.60-2.69 (m, 1H), 2.03-2.12 (m, 1H).

N-(4-fluorophenyl)-2-oxo-1-(4-(piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-1,2-dihydropyridine-3-carboxamide(83)

Retention time (min)=4.282, method [7], MS (ESI) 433.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.11 (s, 1H), 8.59 (dd, 1H), 7.73 (d, 2H), 7.71-7.75(m, 2H), 7.21-7.27 (m, 2H), 7.00-7.26 (m, 2H), 6.91 (d, 1H), 6.73 (d,1H), 6.58 (t, 1H), 6.39 (t, 1H), 2.90-3.07 (m, 10H), 2.76-2.87 (m, 1H),1.97-2.08 (m, 1H).

N-(4-bromophenyl)-2-oxo-1-(4-(piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-1,2-dihydropyridine-3-carboxamide(84)

Retention time (min)=5.915, method [7], MS (ESI) 493.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.16 (s, 1H), 8.56 (dd, 1H), 7.65 (d, 2H), 7.42 (d,2H), 7.19-7.23 (m, 2H), 6.88 (d, 1H), 6.70 (d, 1H), 6.55 (t, 1H), 6.36(t, 1H), 2.87-3.01 (m, 10H), 2.73-2.84 (m, 1H), 1.94-2.05 (m, 1H).

Example 10 10.1. Synthesis ofN-(4-chlorophenyl)-1-(4-(4-(2-hydroxyethyl)piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(85)

N-(4-chlorophenyl)-2-oxo-1-(4-(piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-1,2-dihydropyridine-3-carboxamide(40 mg, 1.0 eq.), potassium carbonate (100 mg, 8.0 eq) were combined in10 ml dry CH₃CN. The mixture was heated at refluxing for 5 mins. To thesolution was added 2-iodoethanol dropwise. The resulting mixture waskept refluxing overnight. The in-organic solid was removed byfiltration. The filtrate was concentrated in vacuo. The crude mixturewas purified on flash column (0˜5% NH₃ in CH₃CN) in silica gel to getwhite solid 40 mg (yield 91%)N-(4-chlorophenyl)-1-(4-(4-(2-hydroxyethyl)piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide.Retention time (min)=4.967, method [7], MS (ESI) 493.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.18 (s, 1H), 8.59 (dd, 1H), 7.73 (d, 2H), 7.22-7.32(m, 4H), 6.92 (d, 1H), 6.74 (d, 1H), 6.58 (t, 1H), 6.39 (t, 1H), 4.51(m, 1H), 3.66-3.69 (m, 2H), 2.63-3.12 (m, 13H), 1.97-2.08 (m, 1H).

10.2. Synthesis ofN-(4-chlorophenyl)-1-(4-(4-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(86)

The title compound was synthesized from appropriate starting materialsusing the procedures outlined above in Example 10.1., or slightlymodified versions thereof. Retention time (min)=5.652, method [7], MS(ESI) 581.2 (M+H); ¹H NMR (300 MHz, DMSO-d6) δ 12.22 (s, 1H), 9.71 (m,1H), 8.44 (dd, 1H), 7.75 (d, 2H), 7.57-7.60 (m, 1H), 7.41 (d, 2H), 7.21(t, 1H), 6.93 (d, 1H), 6.78 (d, 1H), 6.49-58 (m, 2H), 3.76-3.82 (m, 2H),3.54-3.67 (m, 6H), 3.38-3.51 (m, 8H), 2.85-3.32 (m, 6H), 2.62-2.69 (m,1H), 2.06-2.13 (m, 1H).

Example 11 11.1. Synthesis ofN-(4-chlorophenyl)-1-(4-(4-methylpiperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(87)

To a solution ofN-(4-chlorophenyl)-2-oxo-1-(4-(piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-1,2-dihydropyridine-3-carboxamide(40 mg, 1.0 eq.) in 2 ml 1,2-dichloroethane was added formaldehyde (30mg, 10 eq.). The mixture was stirred at rt. for 5 mins. To the reactionmixture was added NaBH(OAc)₃ (34 mg, 1.8 eq.). The resulting mixture wasstirred at rt. for 30 mins; then diluted with 50 ml CH₂Cl₂, washed with10 ml 2M Na2CO3, water and brine. The organic phase was dried oversodium sulfate, and evaporated in vacuo to afford white solid 39 mg(yield 92%)N-(4-chlorophenyl)-1-(4-(4-methylpiperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide.Retention time (min)=5.063, method [7], MS (ESI) 463.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.19 (s, 1H), 8.59 (dd, 1H), 7.73 (d, 2H), 7.21-7.32(m, 4H), 6.92 (d, 1H), 6.73 (d, 1H), 6.57 (t, 1H), 6.39 (t, 1H),2.90-3.11 (m, 6H), 2.78-2.87 (m, 1H), 2.56-2.68 (m, 4H), 2.37 (s, 3H),2.00-2.08 (m, 1H).

11.2. Synthesis of1-(4-(4-methylpiperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(88)

The title compound was synthesized from appropriate starting materialsusing the procedures outlined above in Example 11.1., or slightlymodified versions thereof.

MS (ESI) 430.2 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.34 (s, 1H), 8.56 (dd,1H), 8.50 (d, 2H), 7.65 (d, 2H), 7.19-7.26 (m, 2H), 6.90 (d, 1H), 6.71(d, 1H), 6.54 (t, 1H), 6.38 (t, 1H), 2.74-3.09 (m, 7H), 2.53-2.67 (m,4H), 1.95-2.06 (m, 1H).

Example 12 12.1. Synthesis of1-(4-(4-acetylpiperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-N-(4-chlorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide(89)

To a solution ofN-(4-chlorophenyl)-2-oxo-1-(4-(piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-1,2-dihydropyridine-3-carboxamide(40 mg, 1.0 eq.) in 2 ml CH₂Cl₂ was added DIEA (0.04 ml, 2.5 eq.) andacetyl chloride (9.1 mg, 1.3 eq.). The resulting mixture was stirred atrt. for 40 min; then diluted with 30 ml CH₂Cl₂, washed with 10 ml 2NKHSO4, water, and brine. The crude mixture was purified on flash columnin silica gel to provide white solid 42 mg (yield 96%)1-(4-(4-acetylpiperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-N-(4-chlorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide.Retention time (min)=7.179, method [7], MS (ESI) 491.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.16 (s, 1H), 8.60 (dd, 1H), 7.73 (d, 2H), 7.31 (d,2H), 7.23-7.28 (m, 2H), 6.90 (d, 1H), 6.78 (d, 1H), 6.61 (m, 1H), 6.40(t, 1H), 3.70-3.84 (m, 2H), 3.61-3.67 (m, 2H), 2.90-3.12 (m, 6H),2.81-2.89 (m, 1H), 2.15 (s, 3H), 2.01-2.10 (m, 1H).

12.2. Additional Compounds

The following compounds were synthesized from appropriate startingmaterials using the procedures outlined above in Example 12.1., orslightly modified versions thereof.

1-(4-(4-acetylpiperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(90)

Retention time (min)=1.528, method [7], MS (ESI) 485.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 13.24 (s, 1H), 8.58 (dd, 1H), 8.50 (d, 2H), 8.06 (d,2H), 7.24-7.36 (m, 2H), 6.89 (d, 1H), 6.75 (d, 1H), 6.54 (m, 1H), 6.46(t, 1H), 3.58-3.84 (m, 4H), 2.93-3.09 (m, 6H), 2.80-2.91 (m, 1H), 2.15(s, 3H), 1.97-2.08 (m, 1H).

N-(4-chlorophenyl)-1-(4-(4-(methylsulfonyl)piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(91)

Retention time (min)=8.145, method [7], MS (ESI) 528.1 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.19 (s, 1H), 8.43 (dd, 1H), 7.75 (d, 2H), 7.60(m, 1H), 7.38 (d, 2H), 7.20 (t, 1H), 6.92 (d, 1H), 6.75 (d, 1H),6.49-6.58 (m, 2H), 3.24-3.27 (m, 4H), 3.03-3.12 (m, 5H), 2.86-2.96 (m,4H), 2.60-2.71 (m, 1H), 2.08-2.12 (m, 1H).

Example 13 Synthesis of tert-butyl2-(2-(2-(2-(4-(1-(3-(4-chlorophenylcarbamoyl)-2-oxopyridin-1(2H)-yl)-2,3-dihydro-1H-inden-4-yl)piperazin-1-yl)-2-oxoethoxy)ethoxy)ethoxy)ethylcarbamate(92)

N-(4-chlorophenyl)-2-oxo-1-(4-(piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-1,2-dihydropyridine-3-carboxamide(80 mg, 1.0 eq.) and DIEA (58 mg, 2.5 eq were dissolved in 3 ml CH₂Cl₂.The mixture was cooled over an ice-water bath. A solution of2-bromoacetyl bromide (43.2 mg, 1.2 eq.) in 2 ml CH₂Cl₂ was addeddropwise. The resulting reaction was stirred at 0° C. for 30 mins; thendiluted with 30 ml CH₂Cl₂, washed with 2×15 ml water, 10 ml 2N KHSO4,and brine. The organic phase was dried over sodium sulfate. Removalsolvent in vacuo afforded brown solid 90 mg (yield 89%)1-(4-(4-(2-bromoacetyl)piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-N-(4-chlorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamidewhich was used without further purification.

NaH (60% dispersion in mineral oil) (12.6 mg, 2.0 eq.) was suspended in2 ml dry DMF. A solution of tert-butyl2-(2-(2-hydroxyethoxy)ethoxy)ethylcarbamate (59 mg, 1.5 eq.) in 1 ml DMFwas added dropwise at room temperature. The mixture was heated at 60° C.for 5 mins. The reaction was cooled to room temperature; then a solutionof1-(4-(4-(2-bromoacetyl)piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-N-(4-chlorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide(90 mg, 1.0 eq.) in 1 ml DMF was added. The resulting reaction mixturewas stirred at rt. for 1 hr. The reaction was diluted with 50 ml ethylacetate, washed with 3×20 ml water, and brine. The crude mixture waspurified on HPLC to provide white solid 53 mg (yield 45%) tert-butyl2-(2-(2-(2-(4-(1-(3-(4-chlorophenylcarbamoyl)-2-oxopyridin-1(2H)-yl)-2,3-dihydro-1H-inden-4-yl)piperazin-1-yl)-2-oxoethoxy)ethoxy)ethoxy)ethylcarbamate.Retention time (min)=8.824, method [7], MS (ESI) 760.3 (M+Na); ¹H NMR(300 MHz, CDCl₃) δ 12.15 (s, 1H), 8.60 (dd, 1H), 7.73 (d, 2H), 7.23-7.33(m, 4H), 6.89 (d, 1H), 6.78 (d, 1H), 6.60 (m, 1H), 6.41 (t, 1H), 5.04(m, 1H), 4.27 (s, 2H), 3.60-3.84 (m, 12H), 3.50-3.59 (m, 2H), 3.27-3.32(m 2H), 2.93-3.09 (m, 6H), 2.80-2.90 (m, 1H), 2.00-2.10 (m, 1H), 1.42(s, 9H).

Example 14 Synthesis of1-(4-(4-(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)acetyl)piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-N-(4-chlorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide(93)

To a solution of tert-butyl2-(2-(2-(2-(4-(1-(3-(4-chlorophenylcarbamoyl)-2-oxopyridin-1(2H)-yl)-2,3-dihydro-1H-inden-4-yl)piperazin-1-yl)-2-oxoethoxy)ethoxy)-ethoxy)ethylcarbamate(45 mg) in 2 ml CH₂Cl₂ was added 0.4 ml TFA. The mixture was stirred atroom temperature for 2 hrs. The solvent and TFA were removed in vacuo.The residue was dissolved in 25 ml ethyl acetate, washed with 2×10 ml 2M Na₂CO₃ and brine. The organic phase was dried over sodium sulfate;then evaporated in vacuo to provide white solid 36 mg (yield 93%)1-(4-(4-(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)acetyl)-piperazin-1-yl)-2,3-dihydro-1H-inden-1-yl)-N-(4-chlorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide.Retention time (min)=5.677, method [7], MS (ESI) 639.3 (M+Na); ¹H NMR(300 MHz, DMSO-d6) δ 12.23 (s, 1H), 8.42 (dd, 1H), 7.73 (d, 2H),7.59-7.62 (m, 1H), 7.39 (d, 2H), 7.18 (t, 1H), 6.87 (d, 1H), 6.72 (d,1H), 6.49-6.58 (m, 2H), 4.18 (s, 2H), 3.44-3.56 (m, 6H), 3.29-3.37 (m,4H), 3.27-3.32 (m 2H), 2.92-3.09 (m, 6H), 2.46-2.70 (m, 3H), 2.05-2.14(m, 1H).

Example 15 15.1. Synthesis ofN-(4′-hydroxybiphenyl-4-yl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(94)

N-(4-bromophenyl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(150 mg, 1.0 eq.), Pd(PPh₃)₄ (59 mg, 0.15 eq.) and4-hydroxyphenylboronic acid (61.2 mg, 1.3 eq.) were dissolved in 3 mlDMF. To the solution was added 0.5 m12N Na₂CO₃. The resulting mixturewas heated at 100° C. under N₂ for 30 mins. The reaction was dilutedwith 50 ml ethyl acetate, washed with 3×20 ml water and brine. The crudemixture was purified on flash column in silica gel to afford white solid132 mg (yield 85%)N-(4′-hydroxybiphenyl-4-yl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide.Retention time (min)=8.029, method [7], MS (ESI) 453.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.15 (s, 1H), 8.58 (dd, 1H), 7.80 (d, 2H), 7.44-7.51(m, 4H), 7.16-7.26 (m, 2H), 6.80-6.87 (m, 3H), 6.70 (d, 1H), 6.60 (m,1H), 6.36 (t, 1H), 4.89 (s, 3H), 3.86 (s, 3H), 2.89-3.09 (m, 21H),2.77-2.84 (m, 1H), 1.94-2.05 (m, 1H).

15.2. Additional Compounds

The following compounds were synthesized from appropriate startingmaterials using the procedures outlined above in Example 15.1., orslightly modified versions thereof.

1-(4-Methoxy-2,3-dihydro-1H-inden-1-yl)-N-(4′-methoxybiphenyl-4-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(95)

Retention time (min)=10.232, method [7], MS (ESI) 467.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.12 (s, 1H), 8.58 (dd, 1H), 7.80 (d, 2H), 7.49-7.53(m, 4H), 7.16-7.26 (m, 2H), 6.92 (m, 2H), 6.82 (d, 1H), 6.70 (d, 1H),6.60 (m, 1H), 6.36 (t, 1H), 3.86 (s, 3H), 3.81 (s, 3H), 2.91-3.09 (m,2H), 2.77-2.89 (m, 1H), 1.94-2.05 (m, 1H).

N-(3′-ethoxybiphenyl-4-yl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(96)

Retention time (min)=11.100, method [7], MS (ESI) 481.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.16 (s, 1H), 8.58 (dd, 1H), 7.82 (d, 2H), 7.55 (d,2H), 7.06-7.29 (m, 5H), 6.82 (m, 2H), 6.70 (d, 1H), 6.60 (m, 1H), 6.36(t, 1H), 4.06 (q, 2H), 3.85 (s, 3H), 2.91-3.07 (m, 2H), 2.79-2.89 (m,1H), 1.96-2.05 (m, 1H), 1.41 (t, 3H).

1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-N-(2′-methoxybiphenyl-4-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(97)

Retention time (min)=10.286, method [7], MS (ESI) 467.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.16 (s, 1H), 8.64 (dd, 1H), 7.85 (d, 2H), 7.56 (d,2H), 7.22-7.38 (m, 4H), 6.99-7.07 (m, 2H), 6.78 (d, 1H), 6.76 (d, 1H),6.66 (m, 1H), 6.41 (t, 1H), 3.91 (s, 3H), 3.84 (s, 3H), 2.92-3.15 (m,2H), 2.83-2.90 (m, 1H), 2.00-2.11 (m, 1H).

N-(4′-aminobiphenyl-4-yl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(98)

Retention time (min)=5.056, method [7], MS (ESI) 452.2 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.16 (s, 1H), 8.63 (dd, 1H), 7.83 (d, 2H), 7.55 (d,2H), 7.44 (d, 2H), 7.21-7.32 (m, 2H), 6.87 (d, 1H), 6.75-6.79 (m, 3H),6.66 (m, 1H), 6.41 (t, 1H), 4.89 (s, 3H), 3.91 (s, 3H), 3.73 (s, 2H),2.94-3.13 (m, 2H), 2.82-2.92 (m, 1H), 1.99-2.10 (m, 1H).

Example 16 16.1. Synthesis ofN-(4′-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)biphenyl-4-yl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(99) Protocol K

To a solution ofN-(4′-hydroxybiphenyl-4-yl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(100 mg, 1.0 eq.) in 4 ml dry THF was added tert-butyl2-(2-(2-hydroxyethoxy)ethoxy)ethylcarbamate (60.6 mg, 1.1 eq.), Ph₃P(63.8 mg, 1.1 eq.) and DEAD (42.3 mg, 1.1 eq.). The resulting mixturewas stirred at rt. for overnight. The solvent was removed in vacuo, theresidue mixture was directly purified on flash column in silica gel toprovide white solid 50 mg (yield 32%) tert-butyl2-(2-(2-(4′-(1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamido)biphenyl-4-yloxy)ethoxy)ethoxy)ethylcarbamate.

Protocol L

To a solution of tert-butyl2-(2-(2-(4′-(1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamido)biphenyl-4-yloxy)ethoxy)ethoxy)ethylcarbamate(50 mg) in 2 ml CH₂Cl₂ was added 0.4 ml TFA. The mixture was stirred atroom temperature for 2 hrs. The solvent and TFA were removed in vacuo.The residue was dissolved in 25 ml ethyl acetate, washed with 2×10 ml 2M Na2CO3 and brine. The organic phase was dried over sodium sulfate;then evaporated in vacuo to provide white solid 35 mg (yield 80%)N-(4′-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)biphenyl-4-yl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide.Retention time (min)=6.041, method [7],

MS (ESI) 584.2 (M+H); ¹H NMR (300 MHz, DMSO-d6) δ 12.21 (s, 1H), 8.44(dd, 1H), 7.75-7.82 (m, 4H), 7.54-7.62 (m, 5H), 7.23 (t, 1H), 6.92-7.01(m, 2H), 6.71 (d, 1H), 6.51-6.57 (m, 2H), 4.10-4.13 (m, 2H), 3.81 (s,3H), 3.73-3.76 (m, 2H), 3.57-3.62 (m, 6H), 2.99-3.09 (m, 1H), 2.80-2.96(m, 3H), 2.65-2.75 (m, 1H), 2.02-2.10 (m, 1H).

16.2. Additional Compounds

The following compounds were synthesized from appropriate startingmaterials using the procedures outlined above in Example 16.1., orslightly modified versions thereof.

N-(3′-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)biphenyl-4-yl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(100)

Retention time (min)=6.291, method [7], MS (ESI) 584.3 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.24 (s, 1H), 8.44 (dd, 1H), 7.78-7.82 (m, 4H),7.67 (d, 2H), 7.55-7.58 (m, 1H), 7.34 (t, 1H), 6.88-6.95 (m, 2H), 6.71(d, 1H), 6.52-6.58 (m, 2H), 4.15-4.18 (m, 2H), 3.81 (s, 3H), 3.74-3.77(m, 2H), 3.57-3.63 (m, 6H), 2.99-3.09 (m, 1H), 2.92-2.95 (m, 2H),2.81-2.98 (m, 1H), 2.65-2.75 (m, 1H), 2.00-2.12 (m, 1H).

N-(3′-hydroxybiphenyl-4-yl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(101)

Retention time (min)=8.271, method [7], MS (ESI) 453.2 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.22 (s, 1H), 9.48 (s, 1H), 8.43 (dd, 1H), 7.78(d, 2H), 7.55-7.60 (m, 3H), 7.19-7.25 (m, 2H), 6.92-7.06 (m, 3H), 6.72(d, 1H), 6.55 (m, 1H), 3.81 (s, 3H), 2.91-3.09 (m, 1H), 2.71-2.86 (m,1H), 2.63-2.69 (m, 1H), 2.01-2.12 (m, 1H).

Example 17 Synthesis of tert-butyl2-(2-(2-(4-(4-(1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamido)phenoxy)phenoxy)ethoxy)ethoxy)-ethylcarbamate(102)

1-Fluoro-4-nitrobenzene (692 mg, 1.0 eq), hydroquinone (540 mg, 1.0 eq)and potassium carbonate (2.03 g, 3.0 eq) were combined in 20 ml dry DMF.The mixture was heated at 120° C. for 4 hrs. The in-organic salt wasremoved by filtration. The filtrate was diluted with 200 ml ethylacetate, washed with 3×80 ml water, and brine. The crude mixture waspurified on flash column to give brown solid 210 mg (yield 18%)4-(4-nitrophenoxy) phenol.

To a solution of 4-(4-nitrophenoxy)phenol (180 mg, 1.0 eq.) in 8 ml dryTHF was added tert-butyl 2-(2-(2-hydroxyethoxy)ethoxy)ethylcarbamate(210 mg, 1.1 eq.), Ph₃P (408 mg, 2.0 eq.) and DEAD (271 mg, 2.0 eq.).The resulting mixture was stirred at rt. for overnight. The solvent wasremoved in vacuo, the residue mixture was directly purified on flashcolumn in silica gel to provide yellow solid 340 mg (yield 94%)tert-butyl2-(2-(2-(4-(4-nitrophenoxy)phenoxy)ethoxy)ethoxy)ethylcarbamate.

tert-butyl2-(2-(2-(4-(4-nitrophenoxy)phenoxy)ethoxy)ethoxy)ethylcarbamate (388 mg)was dissolved in 40 ml MeOH, added catalyst (40 mg, 10% Palladium oncarbon). The mixture was hydrogenated (50 psi H₂) at ambient temperaturefor 30 mins. The catalyst was removed by filtration. The solvent wasevaporated in vacuo to give yellow oil 240 mg (yield 75%) tert-butyl2-(2-(2-(4-(4-aminophenoxy)phenoxy)ethoxy)-ethoxy)ethylcarbamate.

Following Protocol C,1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxylicacid was converted to tert-butyl2-(2-(2-(4-(4-(1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamido)phenoxy)phenoxy)ethoxy)ethoxy)ethylcarbamate.Retention time (min)=10.484, method [7], MS (ESI) 722.3 (M+Na); ¹H NMR(300 MHz, CDCl₃) δ 12.02 (s, 1H), 8.56 (dd, 1H), 7.68 (d, 2H), 7.15-7.25(m, 2H), 6.70-6.94 (m, 7H), 6.69 (d, 1H), 6.58 (m, 1H), 6.34 (t, 1H),4.99 (m, 1H), 4.07-4.10 (m, 2H), 3.85 (s, 3H), 3.80-3.84 (m, 2H),3.67-3.70 (m, 2H), 3.60-3.63 (m, 2H), 3.50-3.53 (m, 2H), 3.26-3.31 (m,2H), 2.90-3.06 (m, 2H), 2.75-2.88 (m, 1H), 1.93-2.04 (m, 1H), 1.39 (s,9H).

Example 18 Synthesis ofN-(4-(4-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)phenoxy)phenyl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(103)

To a solution of tert-butyl2-(2-(2-(4-(4-(1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamido)phenoxy)phenoxy)ethoxy)ethoxy)ethylcarbamate (15 mg) in 0.5 ml CH₂Cl₂was added 0.1 ml TFA. The mixture was stirred at room temperature for 2hrs. The solvent and TFA were evaporated in vacuo. The residue wasdissolved in 10 ml ethyl acetate, washed with 2×3 m12 M Na₂CO₃ andbrine. The organic phase was dried over sodium sulfate; then evaporatedin vacuo to provide white solid 9.8 mg (yield 72%)N-(4-(4-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)phenoxy)phenyl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide.Retention time (min)=6.236, method [7], MS (ESI) 600.3 (M+H); ¹H NMR(300 MHz, DMSO-d6) δ 12.11 (s, 1H), 8.41 (dd, 1H), 7.68 (d, 2H), 7.53(m, 1H), 7.22 (d, 1H), 6.90-6.99 (m, 7H), 6.69 (d, 1H), 6.49-6.56 (m,2H), 4.04-4.07 (m, 2H), 3.81 (s, 3H), 3.71-3.74 (m, 2H), 3.56-3.62 (m,6H), 2.98-3.08 (m, 1H), 2.81-2.95 (m, 2H), 2.66-2.79 (m, 1H), 2.62-2.74(m, 1H), 1.99-2.10 (m, 1H).

Example 19 Synthesis of2-oxo-1-(4-(pyridin-3-yl)-2,3-dihydro-1H-inden-1-yl)-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(104)

2-oxo-1-(4-(pyridin-3-yl)-2,3-dihydro-1H-inden-1-yl)-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamidewas prepared by heating a mixture of1-(4-bromo-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(35.1 mg, 0.0855 mmol, 1.0 eq.), pyridin-3-ylboronic acid (12.8 mg,0.104 mmol, 1.2 eq.), Pd(PPh₃)₄ (5.6 mg, 0.0048 mmol, 0.056 eq.), andpotassium carbonate (15.1 mg, 0.109 mmol, 1.3 eq.) in 0.80 mL ofdioxanes and 0.20 mL of H₂O in a microwave at 140° C. for 5 min. Thecrude material was filtered through celite and the filtrate was purifiedby reverse phase HPLC using the method of 5-35% ACN/H₂O in 40 min. Thepurified material was concentrated and dried under vacuum to give2-oxo-1-(4-(pyridin-3-yl)-2,3-dihydro-1H-inden-1-yl)-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamideas a clear colorless oil. Retention time (min)=8.051, method [8], MS(ESI) 409.2 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 9.19 (m, 1H), 8.90 (m, 1H),8.67 (m, 2H), 8.53 (m, 2H), 8.24 (d, J=7.1 Hz, 2H), 8.06 (m, 1H), 7.53(m, 2H), 7.32 (m, 1H), 6.71 (t, J=7.7 Hz, 1H), 6.62 (t, J=7.1 Hz, 1H),3.20 (m, 3H), 2.95 (m 1H), 2.19 (m, 1H).

Example 20 Synthesis of1-(4-nitro-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(105) Protocol I

To a mixture of 4-nitro-2,3-dihydro-1H-inden-1-one (8.0 g, 1.0 eq) inMeOH (125 mL) was added NaBH₄ (0.6 eq) in portions, the mixture wasstirred at room temperature for 40 minutes. At the end of reaction, themixture was concentrated and water was added. The solution was extractedwith ethyl acetate three times. The combined organic layer was washedwith brine and dried over Na₂SO₄, filtrated and evaporated to give4-nitro-2,3-dihydro-1H-inden-1-one (7.9 g, 98%).

Protocol J

To a mixture of 2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(8.0 g, 1.0 eq) in dry DMF (30 mL) was added4-nitro-2,3-dihydro-1H-inden-1-one (1.2 eq) and PPh₃(3.0 eq), then addedDIAD (3.0 eq) under N₂ atmosphere. Then the mixture was stirred at roomtemperature overnight. At the end of reaction, water was added andextracted with ethyl acetate. The combined organic layer was washed withbrine and dried over MgSO₄, filtrated and evaporated to give the crudeproduct, which was purified by Flash Column (PE:EtOAc=1:2), concentratedthe crude product, and recrystallized by EtOAc to give1-(4-nitro-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(3.5 g, 21%). MS (ESI) 377.0 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.22 (s,1H), 8.62 (m, 1H), 8.53 (m, 2H), 8.24 (m, 1H), 7.67 (m, 2H), 7.49 (m,2H), 6.75 (m, 1H), 6.50 (m, 1H), 3.71 (m, 1H), 3.57 (m, 1H), 2.97 (m,1H), 2.21 (m, 2H).

Example 21 Synthesis of1-(4-amino-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(106)

To a mixture of1-(4-nitro-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(3.5 g, 1.0 eq) in EtOH (125 mL) was added SnCl₂*2 H₂O (5.0 eq) inportions, the mixture was stirred at 70° C. overnight. At the end ofreaction, the mixture was concentrated and ethyl acetate was added. Theorganic layer was washed with NaHCO₃ aqueous for three times. Thecombined organic layer was washed with brine and dried over MgSO₄,filtered and evaporated to give the crude product, which was purified byHPLC to afford1-(4-amino-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(3 g, 93%). MS (ESI) 347.0 (M+H); ¹H NMR (300 MHz, DMSO-d6) δ□ 12.44 (s,1H), 8.49 (d, J=3.0 Hz, 2H), 8.46 (d, J=4.2 Hz, 1H), 7.72 (d, J=3.0 Hz,2H), 7.57 (d, J=4.2 Hz, 1H), 6.96 (t, J=4.5 Hz, 1H), 6.57-6.61 (m, 2H),6.45 (t, J=4.2 Hz, 1H), 6.31 (d, J=4.2 Hz, 1H), 5.19 (s, 2H), 2.85-2.90(m, 1H), 2.65-2.75 (m, 2H), 2.00-2.05 (m, 1H).

Example 22 Synthesis of1-(4-cyano-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(107)

To a mixture of1-(4-amino-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(200 mg, 1.0 eq) in H₂O (4 mL) and conc. HCl (0.5 mL) was dropped thematerials of NaNO₂ (1.1 eq) in 1 mL of H₂O at 0° C., the mixture wasstirred at room temperature for 30 minutes. Then this mixture was addedto another flask contained CuCN (9.0 eq), KCN (8.8 eq) and 2 mL H₂O at0° C., maintained stirring for 5 h. At the end of reaction, FeSO4aqueous was added to the mixture, filtrated and washed the solidsubstance with DMSO, evaporated the filtrate to give the crude product,which was purified by HPLC to afford1-(4-cyano-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(20 mg, 10%).

MS (ESI) 357.0 (M+H); ¹H NMR (300 MHz, DMSO-d6) δ 12.31 (s, 1H), 8.48(br, 1H), 8.47 (d, J=4.5 Hz, 2H), 7.83 (d, J=4.5 Hz, 1H), 7.79 (bd, 1H),7.72 (bs, 2H), 7.52 (d, J=4.5 Hz, 1H), 7.45 (t, J=4.5 Hz, 1H), 6.61 (m,2H), 3.3 (overlap with DMSO-d6, 1H), 3.10-3.25 (m, 1H), 2.70-2.80 (m,1H), 2.20-2.27 (m, 1H).

Example 23 Synthesis of1-(4-chloro-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(108)

To a mixture of1-(4-amino-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(600 mg, 1.0 eq) in H₂O (4 mL) and conc. HCl (2 mL) was added dropwiseNaNO₂ (1.1 eq) in H₂O (1 mL) at 0° C., the mixture was stirred at roomtemperature for 30 minutes. Then this mixture was added to another flaskcontaining CuCl (1.5 eq), 2 mL H₂O and 1 mL aqueous HCl at 0° C., themixture was stirred for 5 h. At the end of reaction, saturated Na₂CO₃aqueous was added to adjust to pH 12. The aqueous solution was extractedwith ethyl acetate three times. The combined organic layer was washedwith brine and dried over MgSO₄, evaporated the filtrate to give thecrude product, which was purified by Flash Column (DCM:MeOH=30:1) toafford1-(4-chloro-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(350 mg, 55%). MS (ESI) 366.0 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.31 (s,1H), 8.61 (bd, 1H), 8.53 (bs, 2H), 7.69 (bs, 2H), 7.38 (bd, 1H), 7.26(bm, 2H), 7.03 (d, J=6.0 Hz, 2H), 6.69 (bs, 1H), 6.46 (bs, 1H),3.05-3.25 (bm, 2H), 2.91 (bd, 1H), 2.08 (bd, 1H).

Example 24 Synthesis of1-(4-(methylamino)-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(109) and1-(4-(dimethylamino)-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(110)

To a mixture of1-(4-amino-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(400 mg, 1.0 eq) in DCM (12 mL) and 2 mL of acetic acid was addedNa(OAc)₃BH (3.0 eq) in portions and polymeric formaldehyde (10 eq), themixture was stirred at room temperature overnight. At the end ofreaction, the mixture was concentrated and water was added thenextracted with ethyl acetate. The combined organic layer was washed withbrine and dried over MgSO₄, filtrated and evaporated to give the crudeproduct, which was purified by HPLC to afford1-(4-(methylamino)-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(100 mg, 24%) and1-(4-(dimethylamino)-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(85 mg, 20%).

( ): MS (ESI) 361.0 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.43 (s, 1H), 8.58(d, J=5.4 Hz, 1H), 8.53 (d, J=3.6 Hz, 2H), 7.71 (d, J=3.6 Hz, 2H), 7.28(overlap with CDCl₃, 1H), 7.23 (t, J=6.0 Hz, 1H), 6.61 (d, J=6.0 Hz,1H), 6.56 (t, J=4.8 Hz, 1H), 6.50 (d, J=6.0 Hz, 1H), 6.40 (t, J=5.4 Hz,1H), 2.95 (s, 3H), 2.75-2.90 (m, 3H), 2.00-2.05 (m, 1H).

( ): MS (ESI) 375.0 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.43 (s, 1H), 8.60(dd, J=1.2 Hz, 4.2 Hz, 1H), 8.53 (d, J=3.3 Hz, 2H), 7.71 (d, J=3.3 Hz,2H), 7.31 (dd, J=1.2 Hz, 4.2 Hz, 1H), 7.23 (t, J=4.7 Hz, 1H), 6.88 (d,J=4.7 Hz, 1H), 6.68 (d, J=4.7 Hz, 1H), 6.57 (t, J=4.2 Hz, 1H), 6.43 (t,J=4.2 Hz, 1H), 3.00-3.10 (m, 2H), 2.92 (s, 6H), 2.75-2.90 (m, 1H),2.00-2.05 (m, 1H).

Example 25 Synthesis1-(4-acetamido-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(111)

To a mixture of1-(4-amino-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(140 mg, 1.0 eq) in DCM (8 mL) was added acetyl chloride (1.5 eq) andEt₃N (2.0 eq), the mixture was stirred at room temperature overnight. Atthe end of reaction, the mixture was concentrated and water was addedthen extracted with ethyl acetate. The combined organic layer was washedwith brine and dried over MgSO₄, filtrated and evaporated until the aimproduct precipitated, filtrated and washed with some DCM to afford1-(4-acetamido-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(90 mg, 59%).

MS (ESI) 389.1 (M+H); ¹H NMR (300 MHz, DMSO-d₆) δ 13.06 (s, 1H), 9.56(s, 1H), 8.77 (d, J=3.9 Hz, 2H), 8.51 (d, J=4.2 Hz, 1H), 8.27 (d, J=3.9Hz, 2H), 7.72 (m, 2H), 7.23 (t, J=4.5 Hz, 1H), 6.90 (d, J=4.5 Hz, 1H),6.55 (t, J=4.2 Hz, 1H), 6.57 (t, J=4.2 Hz, 1H), 3.05-3.15 (m, 1H),2.90-3.05 (m, 1H), 2.60-2.75 (m, 1H), 2.05-2.15 (m, 1H), 2.11 (s, 3H).

Example 26 Synthesis1-(4-methyl-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(112)

Following protocol I and J, 4-methyl-2,3-dihydro-1H-inden-1-one wasconverted to1-(4-methyl-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide.MS (ESI) 346.0 (M+H); ¹H NMR (300 MHz, DMSO-d6) δ□ 13.01 (s, 1H), 8.73(d, J=3.6 Hz, 2H), 8.50 (d, J=3.9 Hz, 1H), 8.19 (d, J=3.6 Hz, 2H), 7.68(d, J=3.9 Hz, 1H), 7.18 (d, J=2.6 Hz, 2H), 6.97 (t, J=2.6 Hz, 1H), 6.64(t, J=3.9 Hz, 1H), 6.55 (t, J=4.2 Hz, 1H), 3.07-3.25 (m, 1H), 2.85-3.00(m, 1H), 2.65-2.75 (m, 1H), 2.31 (s, 3H), 2.05-2.15 (m, 1H).

Example 26 Synthesis of2-oxo-1-(4-phenyl-2,3-dihydro-1H-inden-1-yl)-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(113)

To a mixture of1-(4-chloro-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(200 mg, 1.0 eq) in DME (4 mL) was added phenylboronic acid (3.5 eq),Pd(PPh₃)₂Cl₂ (0.2 eq) and aqueous Na₂CO₃ (2M, 2 mL). The mixture wasstirred at 120° C. for 30 mins by microwave. At the end of reaction 50mL of H₂O was added and extracted with ethyl acetate. The combinedorganic layer was washed with brine and dried over MgSO₄, evaporated thefiltrate to give the crude product, which was purified by HPLC to afford2-oxo-1-(4-phenyl-2,3-dihydro-1H-inden-1-yl)-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(21 mg, 10%).

MS (ESI) 408.0 (M+H); ¹H NMR (300 MHz, DMSO-d₆) δ□ 12.93 (s, 1H), 8.71(br, 2H), 8.53 (d, J=3.9 Hz, 1H), 8.15 (d, J=3.3 Hz, 2H), 7.84 (d, J=3.9Hz, 1H), 7.57 (d, J=4.5 Hz, 2H), 7.50 (t, J=4.5 Hz, 2H), 7.37-7.43 (m,3H), 7.17 (d, J=4.5 Hz, 1H), 6.61-6.69 (m, 2H), 3.15-3.25 (m, 1H),3.05-3.15 (m, 1H), 2.60-2.75 (m, 1H), 2.50 (s, 6H), 2.10-2.15 (m, 1H).

Example 27 Synthesis of1-(4-(methylcarbamoyl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(114)

To a mixture of1-(4-bromo-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(250 mg, 1.0 eq) in t-BuOH (15 mL) and DMSO (4 mL) was added thematerials of Pd(OAc)₂ (0.15 eq), dppp (0.15 eq) and Et₃N (2.25 eq), themixture was stirred at 95 V for 18 hours under CO (5.0 atmospherepressure) in a high pressure vessel. At the end of reaction, the solventwas evaporated to give the crude product, which was purified by HPLC toafford1-(2-oxo-3-(pyridin-4-ylcarbamoyl)pyridin-1(2H)-yl)-2,3-dihydro-1H-indene-4-carboxylicacid (90 mg, 39%).

To a mixture of1-(2-oxo-3-(pyridin-4-ylcarbamoyl)pyridin-1(2H)-yl)-2,3-dihydro-1H-indene-4-carboxylicacid (45 mg, 1.0 eq) in DCM (4 mL) was added EDC.HCl (1.5 eq), HOBt (1.5eq), Et₃N (3.0 eq) and MeNH2.HCl (1.1 eq) The mixture was stirred atroom temperature overnight. Water was added and the mixture wasextracted with DCM. The combined organic layer was washed with brine anddried over MgSO₄, evaporated the filtrate to give the crude product,which was purified by HPLC to afford1-(4-(methylcarbamoyl)-2,3-dihydro-1H-inden-1-yl)-2-oxo-N-(pyridin-4-yl)-1,2-dihydropyridine-3-carboxamide(18 mg, 39%). 389.2 (M+H); ¹H NMR (300 MHz, DMSO-d₆) δ 12.97 (s, 1H),8.73 (d, J=3.6 Hz, 2H), 8.51 (dd, J=1.2 Hz, 4.2 Hz, 1H), 8.31 (d, J=2.8Hz, 1H), 8.20 (d, J=3.6 Hz, 2H), 7.74 (d, J=4.2 Hz, 1H), 7.60 (d, J=4.2Hz, 1H), 7.35 (t, J=4.2 Hz, 1H), 7.27 (d, J=4.2 Hz, 2H), 6.65 (t, J=4.2Hz, 1H), 6.55 (t, J=4.2 Hz, 1H), 3.37 (overlap with DMSO-d₆, 2H),2.60-2.75 (m, 1H), 2.05-2.15 (m, 1H).

Example 28 28.1. Synthesis ofN-(4-bromophenyl)-1-(4-hydroxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(115)

To a solution ofN-(4-bromophenyl)-1-(4-methoxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(0.4 g, 0.91 mmol) in Ch2Cl2 (2 mL) at 0 C was added BBr3 (1M in CH₂Cl₂,9 mL). The reaction was stirred at 0 C for 1 hr. It was poured into sat.aqueous NaHCO3 (50 mL). The mixture was extracted with EtOAc (3×50 mL).The combined organic layers were dried and concentrated to a solid. Thesolid was washed with MeOH and CH2Cl2 to afford a white solid. It wasfurther purified by prep. HPLC to affordN-(4-bromophenyl)-1-(4-hydroxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(120 mg, 31%). Retention time (min)=7.127, method [7], MS (ESI) 425.0(M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.19 (s, 1H), 8.61 (dd, J=2.1 Hz, 7.0Hz, 1H), 7.68 (d, J=9.0 Hz, 2H), 7.47 (d, J=9.0 Hz, 2H), 7.15-7.30(overlap with CDCl₃, 2H), 6.81 (d, J=7.8 Hz, 1H), 6.72 (d, J=7.2 Hz,1H), 6.60 (dd, J=5.4 Hz, 7.2 Hz, 1H), 6.41 (t, J=7.0 Hz, 1H), 5.0 (br,1H), 2.80-3.10 (m, 3H), 1.98-2.15 (m, 1H).

28.2. Synthesis ofN-(biphenyl-4-yl)-1-(4-hydroxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(116)

The title compound was synthesized from appropriate starting materialsusing the procedures outlined above in Example 28.1., or slightlymodified versions thereof. Retention time (min)=8.181, method [7], MS(ESI) 423.1 (M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.30 (s, 1H), 8.65 (dd,J=2.1 Hz, 6.9 Hz, 1H), 7.88 (m, 2H), 7.63 (m, 4H), 7.48 (m, 2H), 7.36(m, 1H), 7.26 (dd, J=2.1 Hz, 6.9 Hz, 1H), 7.18 (t, J=7.8 Hz, 1H), 6.89(d, J=7.8 Hz, 1H), 6.72 (d, J=7.8 Hz, 1H), 6.64 (dd, J=5.4 Hz, 8.1 Hz,1H), 6.42 (t, J=6.9 Hz, 1H), 2.80-3.20 (m, 3H), 2.00-2.15 (m, 1H).

Example 29 29.1. Synthesis ofN-(4-bromophenyl)-1-(4-(2-hydroxyethoxy)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(117)

To a solution ofN-(4-bromophenyl)-1-(4-hydroxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(30 mg, 0.071 mmol) in DMF (0.2 mL) was added K2CO3 (30 mg, 0.22 mmol)and 2-chloroethanol (9.3 uL, 0.14 mmol). The reaction mixture was heatedat 70 C overnight. It was filtered and purified by prep HPLC to affordN-(4-bromophenyl)-1-(4-(2-hydroxyethoxy)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(20 mg, 60%). Retention time (min)=7.588, method [7], MS (ESI) 469.1(M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.20 (s, 1H), 8.57 (dd, J=2.1 Hz, 6.9Hz, 1H), 7.65 (d, J=9.0 Hz, 2H), 7.44 (d, J=9.0 Hz, 2H), 7.15-7.30(overlap with CDCl₃, 2H), 6.83 (d, J=8.1 Hz, 1H), 6.73 (d, J=7.5 Hz,1H), 6.58 (dd, J=5.4 Hz, 8.4 Hz, 1H), 6.39 (t, J=6.9 Hz, 1H), 4.14 (m,2H), 4.00 (m, 2H), 2.78-3.15 (m, 3H), 1.90-2.10 (m, 1H).

29.2. Additional Compounds

The following compounds were synthesized from appropriate startingmaterials using the procedures outlined above in Example 29.1., orslightly modified versions thereof.

N-(4-bromophenyl)-1-(4-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(118)

Retention time (min)=7.579, method [7], MS (ESI) 557.1 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.22 (s, 1H), 8.60 (dd, J=1.8 Hz, 7.2 Hz, 1H), 7.71(d, J=9.0 Hz, 2H), 7.48 (d, J=9.0 Hz, 2H), 7.20-7.30 (overlap withCDCl₃, 2H), 6.86 (d, J=8.1 Hz, 1H), 6.75 (d, J=7.5 Hz, 1H), 6.62 (dd,J=5.7 Hz, 7.8 Hz, 1H), 6.42 (t, J=7.2 Hz, 1H), 4.23 (m, 2H), 3.93 (dd,J=4.2 Hz, 8.4 Hz, 2H), 3.60-3.90 (m, 9H), 2.78-3.20 (m, 3H), 1.90-2.10(m, 1H).

N-(biphenyl-4-yl)-1-(4-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamide(119)

Retention time (min)=8.387, method [7], MS (ESI) 555.3 (M+H); ¹H NMR(300 MHz, CDCl₃) δ 12.18 (s, 1H), 8.59 (dd, J=2.1 Hz, 7.2 Hz, 1H), 7.85(m, 2H), 7.58 (m, 4H), 7.41 (m, 2H), 7.17-7.33 (overlap with CDCl₃, 3H),6.82 (d, J=7.8 Hz, 1H), 6.72 (d, J=7.8 Hz, 1H), 6.61 (dd, J=5.7 Hz, 7.8Hz, 1H), 6.38 (t, J=7.2 Hz, 1H), 4.17-4.23 (m, 2H), 3.89 (m, 2H),3.66-3.78 (m, 6H), 3.58-3.65 (m, 2H), 2.78-3.16 (m, 3H), 1.95-2.08 (m,1H).

Example 30 Synthesis of tert-butyl2-(2-(2-(1-(3-(biphenyl-4-ylcarbamoyl)-2-oxopyridin-1(2H)-yl)-2,3-dihydro-1H-inden-4-yloxy)ethoxy)ethoxy)ethylcarbamate(120)

Following Protocol K,N-(biphenyl-4-yl)-1-(4-hydroxy-2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2-dihydropyridine-3-carboxamidewas converted to tert-butyl2-(2-(2-(1-(3-(biphenyl-4-ylcarbamoyl)-2-oxopyridin-1(2H)-yl)-2,3-dihydro-1H-inden-4-yloxy)ethoxy)ethoxy)ethylcarbamate.Retention time (min)=10.622, method [7],

MS (ESI) 676.4 (M+Na); ¹H NMR (300 MHz, CDCl₃) δ 12.33 (s, 1H), 8.64(dd, J=2.1 Hz, 7.2 Hz, 1H), 7.86 (m, 2H), 7.64 (m, 4H), 7.47 (m, 2H),7.20-7.41 (overlap with CDCl₃, 3H), 6.87 (d, J=8.1 Hz, 1H), 6.77 (d,J=7.5 Hz, 1H), 6.64 (dd, J=5.4 Hz, 8.1 Hz, 1H), 6.45 (t, J=7.2 Hz, 1H),4.20-4.30 (m, 2H), 3.94 (t, J=5.1 Hz, 2H), 3.89 (m, 2H), 3.66-3.78 (m,6H), 3.58-3.65 (m, 2H), 2.78-3.16 (m, 3H), 1.95-2.08 (m, 1H), 1.79 (s,9H).

Example 31 Synthesis of1-(4-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)-2,3-dihydro-1H-inden-1-yl)-N-(biphenyl-4-yl)-2-oxo-1,2-dihydropyridine-3-carboxamidetrifluoroacetate (121)

Following Protocol L, tert-butyl2-(2-(2-(1-(3-(biphenyl-4-ylcarbamoyl)-2-oxopyridin-1(2H)-yl)-2,3-dihydro-1H-inden-4-yloxy)ethoxy)ethoxy)ethylcarbamatewas converted to1-(4-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)-2,3-dihydro-1H-inden-1-yl)-N-(biphenyl-4-yl)-2-oxo-1,2-dihydropyridine-3-carboxamidetrifluoroacetate. Retention time (min)=7.005, method [7], MS (ESI) 554.3(M+H); ¹H NMR (300 MHz, CDCl₃) δ 12.23 (s, 1H), 8.58 (dd, J=1.5 Hz, 8.4Hz, 1H), 7.86 (d, J=8.2 Hz, 2H), 7.59 (m, 4H), 7.41 (t, J=7.5 Hz, 2H),7.18-7.33 (overlap with CDCl₃, 3H), 6.80 (d, J=8.1 Hz, 1H), 6.72 (d,J=7.8 Hz, 1H), 6.58 (dd, J=6.6 Hz, 9.6 Hz, 1H), 6.40 (t, J=8.4 Hz, 1H),4.18 (br, 2H), 3.60-3.95 (br, 11H), 3.19 (br, 2H), 2.75-3.15 (m, 3H),1.95-2.08 (m, 1H).

Example 32 In Vitro α4β7 Integrin Adhesion Assay (8866 Cell Adhesion toMadCAM-Fc)

Compounds of the present invention were tested for their ability tocompete with a natural ligand (MadCAM) for binding to α₄β₇ integrin(i.e., block MadCAM-α4/β7 integrin interactions) using the followingcell-based assay.

Materials

H/S++ buffer: 140 mM NaCl, 20 mM HEPES, 1 mM CaCl₂, 1 mM MgCl₂)Assay buffer: H/S++ buffer with 0.3% BSA8866 media recipe (1 Liter): RPMI 1640 Medium (Gibco) (880 ml), 10% FBSfrom (100 ml), Penn/Strep (media aliquots) (10 ml), L-Glutamine (mediaaliquots) (10 ml)

A. Cell Preparation, Cell Harvest and Labeling

8866 cells were split from a carrying flask the day before the assay andincubated in 8866 media at 37° C. and 5% CO₂.

Harvest six flasks of 8866 cells into two 500 ml conical tubes. Spindown for 5 minutes at 1000 rpm. Aspirate the supernatants and re-suspendthe cells in assay buffer. Wash the cells with 50 ml of assay buffer,gently mix by pipetting up and down several times, transfer to 50 mltube to be centrifuged again for 5 minutes at 1000 rpm.

Aspirate the wash and re-suspend the cells in 16 ml of assay buffer. Mixgently with pipette (carefully w/o bubbles) and then add 16 μl ofCalcein AM (1:1000 10 mg/ml; Invitrogen). Invert gently several timesand then incubate with agitation for 30 min at RT while covering thetube with foil (calcein AM is light sensitive). After incubation, addassay buffer to 50 ml, mix well and count the cells. After counting,spin down the cells at 1000 rpm and re-suspend at 2.5×10⁶ cells/ml inassay buffer.

B. MadCAM Plate Coating

Prepare Mouse ascites anti Hu Fc (Sigma 1-6260) at 1:300. Add 180 μl ofstock into 54 ml of H/S⁺⁺ buffer. Add 100 μl/well to 96-well ELISA plate(Costar 3590) and incubate at RT for 1 hour.

Aspirate the plate, blot, and block the plate with 100 μl/well assaybuffer at RT for 1 hour.

Prepare MadCAM solution at 0.15 μg/ml-add 14.7 μl of MadCAM-1/Fc stockat 0.55 mg/ml into 54 ml of PBS⁺⁺.

Aspirate the plates, wash 3 times with 150 μl/well assay buffer, add 100μl/well of respective MadCAM-1/Fc to the plates and incubate overnightat 4° C.

Wash the MadCAM-Fc coated plates 3 times with 150 μl/well of assaybuffer. Leave the last wash in until ready to add cells/compounds toMadCAM-Fc plates.

C. Preparation of Compound Solutions (a) Reference Compound (ELAN91852-8)

Prepare 604.08 μM by adding 42.34 of 10 mM (DMSO) stock into 0.7 mL ofmedia. Add this compound to A1-A3 of 100% plates and use the Hamiltonprogram “α4β7 1:3” to dilute compound 1 to 3.

(b) Test Compounds

For a 10 μM starting concentration, dilute 15 μL of compound with 285 μLof DMSO. Transfer into triplicates of row A of 100% plates and dilute 1to 3 using the Hamilton. For 100 μM starting conc., dilute 105 μLcompound with 105 μL of DMSO and add triplicates to 100% plates to bediluted 1:3.

Use the Multimek program “147tranf” to transfer 147 μl of assay bufferto each well (w/ exception of row H) of the 2% plates. Then use themultimek program “3 μltrans” to transfer 3 μl of compound from the 100%plates to add to the 147 μl of buffer in the 2% plates. Keep allcompound plates on shaker when not in use.

D. Preparation of 21/6—Positive Control

Prepare 21/6 at a working conc. of 20 μg/ml for a final conc. of 10μg/ml. Add 20 μg/ml solution of 21/6 to wells H4-H12 of all the 2%plates.

E. Assay

Bring the cells in the deep well, the compounds in the 2% plates, andempty 1% plates to the multimek. Transfer 70 μl/well of cells from deepwell plate to 1% plates followed by adding 70 μl/well of the respectivecompounds from 2% plates to 1% plates. (No cells or compounds in H1-H3).Incubate cells+compounds for 30 min. at RT.

Then transfer 100 μl/well from 1% plates to MadCAM-Fc plates. Mix welland consistently in order to have enough cells in the wells. Incubatefor another 30 min at RT.

Carefully wash the plates 4 times with 100 μl/well. Aspirate wells H1-H3after plate is ready and add 50 μl media+50 μl 2× cells=total input.

Read the plates using Cytoflour (Excitation=485, Emission=530, andGain=47).

Example 33 In Vitro Biological Evaluation (SRU MadCAM Assay)

Compounds of the present invention were tested for their ability tocompete with a natural ligand (MadCAM) for binding to α₄β₇ integrinusing the following assay. The SRU Bind platform is a label free assayplatform that consists of three parts: (1) a plate-based opticalbiosensor (2) the Bind Reader, and (3) a software analysis packet. Thissystem offers advantages over manual methods in terms of increasedthroughput, greater sensitivity, and lower assay to assay variability.Specifically, this assay measures adhesion of 8866 cells endogenouslyexpressing a4b7 to immobilized recombinant and soluble MadCAM, which wasexpressed as a fusion protein with a human IgGFc tail.

The SRU Bind cell adhesion assay is a two-day assay. On the first day,the 384-well SRU Bind biosensor plate was hydrated for 15 minutes,washed with Hepes buffer pH 7.4 containing 100 mM NaCl and thenincubated with a monoclonal antibody to human Fc (Sigma 1-6260 1:500dilution in Hepes pH 7.4 buffer) for 60 minutes at room temperature. Theplate was then blocked with 4% BSA in 20 mM Hepes buffer pH 7.4, for onehour and then incubated overnight with 0.15 μg/ml of recombinant MadCAMat 4° C. On the second day, test compounds were serially diluted 1:2 inDMSO (200× final concentration) for a 10-point dose response curve.Compounds were then diluted 50× in 20 mM Hepes buffer pH 7.4 containing1 mM MgCl₂, 1 mM CaCl₂, 140 mM NaCl, and 0.3% BSA (Buffer A) using aMultimek liquid handler. 8866 cells grown in 500 ml conical tubes wereharvested (4×-final) in Buffer A and were incubated with the testcompounds (1:1) for thirty minutes at room temperature. The biosensorassay plate was washed with Buffer A containing 1% DMSO leaving 25 μlin-wells and a baseline measurement was taken. 25 μl of 8866 cells thathad been incubated with compound were then transferred to the SRU Bindassay plate (50000 cells/well final at a 1% DMSO final concentration)and the extent of cell adhesion was measured at 45 minutes. The abilityof compounds to block a4b7-mediated 8866 cell adhesion was thenquantified and plotted using Excel-fit.

Example 34 Cell Adhesion Assay

Compounds of this invention can be tested for their ability to inhibitcellular adhesion. Using RPMI-8866 cells, adhesion to recombinant,immobilized soluble MadCAM-1 can be measured. This assay is described byTidswell et al., J. Immunol. (1997) 159(3):1497-1505.

Example 35 Soluble MadCAM-1 FACS Assay

This assay measures the interaction of recombinant soluble MadCAM-1 withRPMI-8866 cells in suspension. Recombinant soluble MadCAM-1(“rsMadCAM-1”) was expressed as a fusion protein with a human IgG Fctail (Tidswell et al., J. Immunol. (1997) 159(3):1497-1505). SolubleMadCAM-1 was mixed with RPMI-8866 cells in the presence and absence oftest compounds. 1 mM MnCl₂ was included in the assay buffer to increasethe activity of α₄β₇ integrin and to promote its interaction with theMadCAM-1 construct. After 30 minutes at room temperature, the cells werewashed with buffer containing 1 mM MnCl₂, and were exposed to afluorescently labeled antibody against the Fc tail of the MadCAM-1fusion protein in the presence of 1 mM MnCl₂, for 30 minutes at 4° C.The cells were washed, re-suspended in MnCl₂ containing buffer andexamined by FACS analysis. An identical assay can be performed tomeasure the interaction of recombinant soluble VCAM-1 with cells thatexpress α₄β₁, such as the Jurkat T cell line.

Example 35 Cell Free ELISA Assay

This assay measures the interaction of solubilized integrin withMadCAM-1 immobilized on a plastic surface. RPMI-8866 cells were lysedwith a detergent to solubilize the α₄β₇ integrin. An antibody against β₇integrin (2G3) was added to the lysate. See Tidswell et al. J. Immunol.(1997) 159(3):1497-1505. This antibody serves two purposes. First, it isa tag by which α₄β₇ integrin can be detected in the assay and, second,2G3 is an antibody that stabilizes a ligand occupied conformation of β₇integrin and promotes β₇ integrin-dependent interactions. Cell lysate,2G3, and test compound were added to microtiter wells coated withMadCAM-1. The mixture was incubated for 30 minutes at room temperature.The plate was washed, blocked with 1% BSA, and exposed to HRP-conjugatedgoat anti-mouse Ig, which recognizes 2G3 associated with MadCAM-boundα₄β₇ integrin. After 30 minutes at room temperature, the wells werewashed and exposed to a substrate for HRP to quantify the amount of α₄β₇integrin bound to MadCAM-1.

Example 36 FACS Assay for Receptor Occupancy

This assay measures the interaction of antibody 2G3 with RPMI-8866 cellsor with lymphocytes. The antibody recognizes a ligand-occupied epitopeof either rat or human β₇ integrin. Increasing concentrations of smallmolecule ligand induce the 2G3 epitope on β₇ integrin and will allowhigher levels of antibody binding to the surface of the cells. Theconcentration of ligand required for receptor occupancy is directlyrelated to the ligand's affinity for α₄β₇ integrin. A similar assay hasbeen described for examining the interaction of ligands with α₄β₁integrin, which utilizes an analogous antibody against a ligand occupiedepitope of β₁ integrin (antibody 15/7; Yednock et al. (1995) JBC270:28740-50). The δ₁ integrin assay relies on cells that express α₄β₁integrin, rather than α₄β₇ integrin (such as Jurkat cells). In bothassays, the appropriate cells are mixed with either 2G3 or 15/7 in thepresence of the small molecule ligand. The cells are incubated at roomtemperature for 30 minutes and washed to remove unbound antibody. Thecells are exposed to a fluorescently-labeled antibody against mouse IgG,which detects cell-associated 2G3 or 15/7 and the cells are examined byFACS analysis.

Example 37 Ex Vivo Cell Adhesion Assay

This assay measures the adhesion of lymphocytes or RPMI-8866 cells tohigh endothelial venules exposed in tissue sections of Peyer's Patches(lymphoid tissue associated with the intestine). These vessels expresshigh levels of MadCAM-1. This assay is described by Yednock et al., JCB(1987) 104:725-731.

Example 38 In Vivo Migration Assay

Migration of In¹¹¹-labeled or fluorescently-labeled lymphocytes toPeyer's Patches in vivo. In this assay, lymphocytes are isolated fromone group of animals and are labeled with a radioactive or fluorescenttracer. The cells are injected intravenously into a second group ofanimals. After 1 to 24 hours, the localization of the labeled cells todifferent tissues can be monitored by either determining the number ofradioactive counts associated with different tissues in a gamma counter,or by isolating lymphocytes from the tissue and determining the numberof cells that carry a fluorescent tag (determined by FACS analysis).This type of assay is described by Rosen et al., J. Immunol. 1989,142:1895-1902.

Example 39 In Vivo Biological Evaluation (Asthma Models)

Inflammatory conditions mediated by α₄β₁ integrin include, for example,eosinophil influx, airway hyper-responsiveness and occlusion that occurswith chronic asthma. The following describes animal models of asthmathat are used to study the in vivo effects of the compounds of thisinvention for use in treating asthma.

39.1. Rat Asthma Model (E2)

Following the procedures described by Chapman, et al., Am J. Resp. Crit.Care Med., 153-4, A219 (1996) and Chapman, et al., Am. J. Resp. Crit.Care Med. 155:4, A881 (1997), both of which are incorporated byreference in their entirety.

Ovalbumin (OA; 10 μg/mL) is mixed with aluminum hydroxide (10 mg/mL) andinjected (i.p.) in Brown Norway rats on day 0. Injections of OA,together with adjuvant, are repeated on days 7 and 14. On day 21,sensitized animals are restrained in plastic tubes and exposed (60minutes) to an aerosol of OA (10 mg/kg) in a nose-only exposure system.Animals are sacrificed 72 hours later with pentobarbital (250 mg/kg,i.p.). The lungs are lavaged via a tracheal cannula using 3 aliquots (4mL) of Hank's solution (HBSS×10, 100 ml; EDTA 100 mM, 100 mL; HEPES 1 M,25 mL; made up to 1 L with H₂O); recovered cells are pooled and thetotal volume of recovered fluid adjusted to 12 mL by addition of Hank'ssolution. Total cells are counted (Sysmex microcell counter F-500, TOAMedical Electronics Otd., Japan) and smears are made by dilutingrecovered fluid (to approximately 106 cells/mL) and pipetting an aliquot(100 μl) into a centrifuge (Cytospin, Shandon, U.K.). Smears are airdried, fixed using a solution of fast green in methanol (2 mg/mL) for 5seconds and stained with eosin G (5 seconds) and thiazine (5 seconds)(Diff-Quick, Browne Ltd. U.K.) in order to differentiate eosinophils,neutrophils, macrophages and lymphocytes. A total of 500 cells per smearare counted by light microscopy under oil immersion (×100). Compounds ofthis invention can be formulated into a 0.5% carboxymethylcellulose and2% Tween 80 suspension and administered orally to rats which had beensensitized to the allergen, ovalbumin. Compounds which inhibitedallergen-induced leukocyte accumulation in the airways of activelysensitized Brown Norway rats are considered to be active in this model.

39.2. Mouse Asthma Model (E3)

Compounds are also evaluated in a mouse model of acute pulmonaryinflammation following the procedures described by, Kung, et al., Am J.Respir. Cell Mol. Biol., 13:360-365, (1995) and Schneider, et al.,(1999). Am J. Respir. Cell Mol. Biol. 20:448-457, (1999), which are eachincorporated by reference in their entirety. Female Black/6 mice (8-12weeks of age) are sensitized on day 1 by an intraperitoneal injection of0.2 mL ova/alum mixture containing 20 μg of ova (Grade 4, Sigma) and 2mg inject Alum (Pierce). A booster injection is administered on day 14.Mice are challenged on days 28 and 29 with aerosolized 1% ova (in 0.9%saline) for 20 minutes. Mice are euthanized and bronchaveolar lavagesamples (3 mL) are collected on day 30, 48 hours post first challenge.Eosinophils are quantified by a FACS/FITC staining method. Compounds ofthis invention are formulated into a 0.5% carboxymethylcellulose and 2%Tween 80 suspension and administered orally to mice which had beensensitized to the allergen, ovalbumin. Compounds which inhibitedallergen-induced leucocyte accumulation in the airways of activelysensitized C57BL/6 mice are considered to be active in this model.

39.3. Sheep Asthma Model (E4)

This model employs the procedures described by Abraham, et al., J. Clin,Invest, 93:776-787 (1994) and Abraham, et al., Am J. Respir. Crit. CareMed., 156:696-703 (1997), both of which are incorporated by reference intheir entirety. Compounds of this invention are evaluated by intravenous(saline aqueous solution), oral (2% Tween 80, 0.5%carboxymethylcellulose), and aerosol administration to sheep which arehypersensitive to Ascaris suum antigen. Compounds which decrease theearly antigen-induced bronchial response and/or block the late-phaseairway response, e.g. have a protective effect against antigen-inducedlate responses and airway hyper-responsiveness (“AHR”), are consideredto be active in this model.

Allergic sheep which are shown to develop both early and late bronchialresponses to inhaled Ascaris suum antigen are used to study the airwayeffects of the candidate compounds. Following topical anesthesia of thenasal passages with 2% lidocaine, a balloon catheter is advanced throughone nostril into the lower esophagus. The animals are then incubatedwith a cuffed endotracheal tube through the other nostril with aflexible fiberoptic bronchoscope as a guide.

Pleural pressure is estimated according to Abraham (1994). Aerosols (seeformulation below) are generated using a disposable medical nebulizerthat provided an aerosol with a mass median aerodynamic diameter of 3.2μm as determined with an Andersen cascade impactor. The nebulizer isconnected to a dosimeter system consisting of a solenoid valve and asource of compressed air (20 psi). The output of the nebulizer isdirected into a plastic T-piece, one end of which is connected to theinspiratory port of a piston respirator. The solenoid valve is activatedfor 1 second at the beginning of the inspiratory cycle of therespirator. Aerosols are delivered at VT of 500 mL and a rate of 20breaths/minute. A 0.5% sodium bicarbonate solution only is used as acontrol.

To assess bronchial responsiveness, cumulative concentration-responsecurves to carbachol is generated according to Abraham (1994). Bronchialbiopsies are taken prior to and following the initiation of treatmentand 24 hours after antigen challenge. Bronchial biopsies are preformedaccording to Abraham (1994).

An in vitro adhesion study of alveolar macrophages can also be performedaccording to Abraham (1994), and a percentage of adherent cells can becalculated.

Aerosol Formulation

A solution of compound n 0.5% sodium bicarbonate/saline (w/v) at aconcentration of 30.0 mg/mL is prepared using the following procedure:

A. Preparation of 0.5% Sodium Bicarbonate/Saline Stock Solution: 100.0mL

Final Ingredient Gram/100.0 mL Concentration Sodium 0.5 g 0.5%Bicarbonate Saline q.s. ad 100.0 mL q.s. ad 100%

Procedure:

-   1. Add 0.5 g sodium bicarbonate into a 100 mL volumetric flask.-   2. Add approximately 90.0 mL saline and sonicate until dissolved.-   3. Q.S. to 100.0 mL with saline and mix thoroughly.    B. Preparation of 30.0 mg/mL Compound: 10.0 mL

Final Ingredient Gram/10.0 mL Concentration Compound 0.300 g 30.0 mg/mL0.5% Sodium q.s. ad 10.0 mL q.s ad 100% Bicarbonate/ Saline StockSolution

Procedure:

-   1. Add 0.300 g of the compound into a 10.0 mL volumetric flask.-   2. Add approximately 9.7 mL of 0.5% sodium bicarbonate/saline stock    solution.-   3. Sonicate until the compound is completely dissolved.-   4. Q.S. to 10.0 mL with 0.5% sodium bicarbonate/saline stock    solution and mix thoroughly.

Example 40 In Vivo Biological Evaluation (Arthritis) 40.1.Adjuvant-Induced Arthritis in Rats

Adjuvant induced arthritis (“AIA”) is an animal model useful in thestudy of rheumatoid arthritis (“RA”), which is induced by injecting M.tuberculosis in the base of the tail of Lewis rats. Between 10 and 15days following injection, animals develop a severe, progressivearthritis.

The compounds of the invention can be tested for their ability to alterhind paw swelling and bone damage resulting from adjuvant induced edemain rats. To quantitate the inhibition of hind paw swelling resultingfrom AIA, two phases of inflammation have been defined: (1) the primaryand secondary injected hind paw, and (2) the secondary non-injected hindpaw, which generally begins developing about eleven days from theinduction of inflammation in the injected paw. Reduction of the lattertype of inflammation is an indication of immunosuppressive activity. C FChang, Arth. Rheum., 20, 1135-1141 (1977).

Using an animal model of RA, such as AIA, enables one to study thecellular events involved in the early stages of the disease. CD44expression on macrophages and lymphocytes is up regulated during theearly development of adjuvant arthritis, whereas LFA 1 expression is upregulated later in the development of the disease. Understanding theinteractions between adhesion molecules and endothelium at the earlieststages of adjuvant arthritis could lead to significant advances in themethods used in the treatment of RA.

40.2. Collagen Induced Arthritis in Rats

Compounds of the invention can be tested in this animal model bymeasuring the inhibition of inflammation, cartilage destruction and boneresorption that occurs in developing type II collagen arthritis in rats.

Animals: 54 Female Lewis rats (Harlan), weighing 125-150 g on arrival.(inject 50 with collagen to get 50 responders on days 10, 11, 12 for 6groups of 10). The animals (10/group for arthritis, 4/group for normalcontrol), housed 4-5/cage, were acclimated for 4-8 days. The animalswere dosed at po3 mg/kg bid, po10 mg/kg bid, and po30 mg/kg bid.

Materials: Agents or drugs in vehicle, Type II collagen, Freund'sincomplete adjuvant, methotrexate (Sigma), compounds of the invention.

General Study Design

Dosing was initiated on day minus 1. The acclimated animals wereanesthetized with isoflurane and given collagen injections (D0). On day6 they were anesthetized again for the second collagen injection.Collagen was prepared by making a 4 mg/mL solution in 0.01N acetic acid.Equal volumes of collagen and Freund's incomplete adjuvant, wereemulsified by hand mixing until a bead of this material held its formwhen placed in water. Each animal received 300 μA of the mixture eachtime spread over 3 sites on back. Calipering of normal (pre-disease)right and left ankle joints were done on day 9. On days 10-12, onset ofarthritis occurred.

Rats were weighed on days (−) 1, 6, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, and 20 of the study and caliper measurements of ankles takenevery day beginning on day 9. Final body weights were taken on day 20.After final body weight measurement, animals were anesthetized forterminal plasma collection and then euthanized.

Both hind paws and knees were removed. Hind paws were weighed, placed(with knees) in formalin and then processed for microscopy.

Processing of Joints

Following 1-2 days in fixative and then 4-5 days in decalcifier, theankle joints were cut in half longitudinally, knees were cut in half inthe frontal plane, processed, embedded, sectioned and stained withtoluidine blue.

Certain compounds of the invention exhibited significant inhibitioncompared to controls receiving no treatment of ankle inflammation andankle histopathology at doses tested (3.0 mg/kg, 10.0 mg/kg and 30.0mg/kg).

Example 41 In Vivo Biological Evaluation (Colitis) Induction of Colitisin HLA-B27 Rats

The efficacy of the compounds of the present invention in reversingcolitis was determined in HLA-B27 transgenic rats. HLA-B27 transgenicrats have been utilized as an animal model of Inflammatory Bowel Diseasewhich mimics Crohn's Disease in humans. The rats overexpress the humanMHC class I HLA-B27 heavy chain and beta-2 microglobulin proteins, whichinduces a variety of autoimmune diseases that include inflammation ofthe colon.

The therapeutic effect of the compounds of this invention in resolvingcolitis was evaluated in HLA-B27 transgenic rats. Diseased rats weredosed subcutaneously with 100 mg/kg of a selected compound of thisinvention twice a day for 16 days. Animal samples dosed with 100 mg/kgof the compound of this invention showed clinical and histologicalresolution of colitis and mimicked similar efficacy with the positivecontrol group treated with anti-alpha4 antibody GG5/3.

Disease Activity Index (DAI) scores indicated overall improved scoresfor rats dosed with 100 mg/kg of certain compounds of this invention and10 mg/kg of GG5/3 (positive control) than rats dosed with vehicle. Fecalconsistency and FOB scores for rats dosed with the compound and GG5/3were statistically different from the vehicle group.

Induction of Colitis

20 HLA-B27 (6-9 weeks old) transgenic rats were ordered from Taconic.Rats acclimated in animal facility for 10 weeks. Animal bedding wasmixed from different cages once a week to control for a “dirty”environmental flora. Certain compounds of the invention inhibit kainicacid-induced phospho-cJun upregulation in Mice.

1.-19. (canceled)
 20. A pharmaceutical composition comprising a compoundof claim 29 and a pharmaceutically acceptable carrier.
 21. A method oftreating an inflammatory disease comprising administering to a mammaliansubject in need thereof a pharmaceutically effective amount of acompound according to claim 29, wherein the inflammatory disease isselected from the group consisting of asthma, inflammatory boweldisease, ulcerative colitis, Crohn's disease, multiple sclerosis,rheumatoid arthritis, tumor metastasis, graft versus host disease, andorgan or tissue rejection.
 22. The method of claim 21, wherein saidinflammatory disease is a member selected from Crohn's disease andulcerative colitis.
 23. An in vitro assay for measuring binding of anα4β1 or α4β7 integrin to an integrin ligand, wherein the assaycomprises: (i) binding the ligand to a surface; (ii) contacting theligand with a cell expressing the integrin, in the presence of acompound of claim 29; and (iii) measuring the amount of cells bound tothe surface.
 24. The assay according to claim 23, wherein the integrinligand is a member selected from fibronectin (FN), VCAM-1, osteopontinand MadCAM.
 25. (canceled)
 26. An in vitro assay for measuring bindingof the compound to an α4β1 or α4β7 integrin in the presence of acandidate molecule, wherein the assay comprises incubating the testmolecule in the presence of a compound of claim 29 labeled with aradioactive, colorimetric or fluorescent label, and measuring the amountof the labeled compound for binding to the integrin.
 27. An in vitroassay for identifying a candidate molecule capable of binding to α4β1 orα4β7 integrin, wherein the assay comprises incubating the candidatemolecule in the presence of a compound of claim 29 labeled with aradioactive, colorimetric or fluorescent label, and measuring the amountof the labeled compound for binding to the integrin, and wherein thecandidate molecule is identified as capable of binding to the integrinif it exhibits a binding activity of an IC₅₀ of not more than 10 μM inthe assay.
 28. (canceled)
 29. A compound of Formula (Ia′), Formula (Ib),or Formula (Ic)

or a salt or solvate or single stereoisomer or mixture of stereoisomersthereof, wherein m is an integer selected from 0 to 4; n is an integerselected from 0 to 3; p is an integer selected from 0 to 4; m1 is aninteger selected from 0 to 3; ring A is a member selected fromsubstituted or unsubstituted aryl and substituted or unsubstitutedheteroaryl; Cy is a member selected from substituted or unsubstituted(C₃-C₁₀)cycloalkyl, substituted or unsubstituted 3- to 10-memberedheterocycloalkyl, substituted or unsubstituted aryl and substituted orunsubstituted heteroaryl; E is selected from O, S, C(O), S(O)₂ and NR⁴⁰,wherein R⁴⁰ is a member selected from substituted or unsubstitutedalkyl; each R¹, each R², R^(1a) and R^(1b) are a member independentlyselected from H, substituted or unsubstituted (C₁-C₁₀)alkyl, substitutedor unsubstituted 2- to 10-membered heteroalkyl, substituted orunsubstituted (C₃-C₁₀)cycloalkyl, substituted or unsubstituted 3- to10-membered heterocycloalkyl, substituted or unsubstituted aryl,substituted or unsubstituted heteroaryl, nitro, CN, halogen, OR¹², SR¹²,NR¹²R¹³, C(O)R¹⁴, C(O)NR¹²R¹³, OC(O)NR¹²R¹³, C(O)OR¹², NR¹⁵C(O)R¹⁴,NR¹⁵C(O)OR¹², NR¹⁵C(O)NR¹²R¹³, NR¹⁵C(S)NR¹²R¹³, NR¹⁵S(O)₂R¹⁴,S(O)₂NR¹²R¹³ and S(O)_(z)R¹⁴, wherein z is 1 or 2; R¹², R¹³ and R¹⁵ aremembers independently selected from H, acyl, substituted orunsubstituted (C₁-C₁₀)alkyl, substituted or unsubstituted 2- to10-membered heteroalkyl, substituted or unsubstituted aryl, substitutedor unsubstituted heteroaryl, substituted or unsubstituted(C₃-C₁₀)cycloalkyl, substituted or unsubstituted 3- to 10-memberedheterocycloalkyl; and R¹⁴ is a member independently selected fromsubstituted or unsubstituted (C₁-C₁₀)alkyl, substituted or unsubstituted2- to 10-membered heteroalkyl, substituted or unsubstituted aryl,substituted or unsubstituted heteroaryl, substituted or unsubstituted(C₃-C₁₀)cycloalkyl, substituted or unsubstituted 3- to 10-memberedheterocycloalkyl, wherein R¹² and R¹³, together with the nitrogen atomsto which they are attached, are optionally joined to form a 4- to7-membered ring, and wherein two adjacent R¹, together with the atoms towhich they are attached, are optionally joined to form a 5- to7-membered ring, and wherein two adjacent R², together with the atoms towhich they are attached, are optionally joined to form a 5- to7-membered ring; R³, R⁴ and R⁵ are members independently selected fromH, substituted or unsubstituted (C₁-C₄)alkyl, halogen and CN; and R⁶ isa member selected from H and substituted or unsubstituted (C₁-C₄)alkyl.30. The compound of claim 29, wherein the compound is of Formula (Ia′):

or a salt or solvate or single stereoisomer or mixture of stereoisomersthereof.
 31. The compound of claim 29, wherein the compound is ofFormula (IIa′):

or a salt or solvate or single stereoisomer or mixture of stereoisomersthereof, wherein m, n, p, Cy, A, R¹, and R² are as defined in claim 29.32. The compound of claim 29, wherein the compound is of Formula(IIIa′):

or a salt or solvate or single stereoisomer or mixture of stereoisomersthereof, wherein X¹, X², X³ and X⁴ are members independently selectedfrom N and CR², q is an integer selected from 0 to 4, and m, Cy, R¹, R²,R³, R⁴, R⁵ and R⁶ are as defined in claim
 29. 33. The compound of claim29, wherein the compound is of Formula (Ib):

or a salt or solvate or single stereoisomer or mixture of stereoisomersthereof.
 34. The compound of claim 29, wherein the compound is ofFormula (IIb):

or a salt or solvate or single stereoisomer or mixture of stereoisomersthereof, wherein Cy, R^(1a), A, R² and p are as defined in claim
 29. 35.The compound of claim 29, wherein the compound is of Formula (Ic):

or a salt or solvate or single stereoisomer or mixture of stereoisomersthereof.
 36. The compound of claim 29, wherein ring A is a memberselected from substituted or unsubstituted phenyl, substituted orunsubstituted pyridyl, substituted or unsubstituted thiazole,substituted or unsubstituted imidazolyl and substituted or unsubstitutedthiophene.
 37. The compound of claim 29, wherein R^(1a) and R^(1b) areH.
 38. The compound of claim 29, wherein R³, R⁴ and R⁵ are eachindependently selected from H and F.
 39. The compound of claim 29,wherein n is 1 or 2, m is 0 or 1, and p is 0 or
 1. 40. The compound ofclaim 29, wherein R⁶ is H.
 41. The compound of claim 29, wherein Cy is amember selected from substituted or unsubstituted phenyl and substitutedor unsubstituted pyridyl.
 42. A compound selected from FIG. 2, or a saltor a solvate thereof.